Pharmacokinetics of S-Adenosylmethionine Formulations

ABSTRACT

Compositions and methods to improve the pharmacokinetic profile of S-Adenosylmethionine (SAMe) are provided, as are methods of treating various disorders using SAMe formulations with improved pharmacokinetic profiles. More specifically, the invention is directed to methods of treating a disease or disorder in a subject and/or improving the nutritional status of a subject by administering formulations exhibiting improved pharmacokinetic profiles of exogenous SAMe. The method also includes the step of orally administering compositions of the invention to the subject once per day after overnight fast; that is prior to food intake in the morning.

CROSS-REFERENCE TO RELATED APPLICATIONS AND CLAIM TO PRIORITY

This application is a divisional application of U.S. patent applicationSer. No. 12/845,600, filed Jul. 28, 2010, now U.S. Pat. No. 8,329,208,issued Dec. 11, 2012, which claims priority to U.S. Provisional patentapplication Ser. No. 61/229,186, filed Jul. 28, 2009, which isincorporated herein by reference in its entirety.

TECHNICAL FIELD

The invention relates to compositions and methods for improving thepharmacokinetic profile of S-adenosyl-L-methionine (“SAM-e” or “SAMe”).More particularly, the invention concerns formulations that lead to SAMeplasma concentrations and AUC values that are increased in comparison tosimilar or higher doses of conventional SAMe formulations. The inventionis directed to methods of treating a disease or disorder in a subjectand/or improving the nutritional status of a subject by administeringformulations exhibiting improved pharmacokinetic profiles of exogenousSAMe. The method also includes orally administering compositions of theinvention to the subject once per day after overnight fast, that isprior to food intake in the morning, which may alleviate some of theside effects (e.g. insomnia and gastrointestinal) associated withconventional twice-daily (or more) dosing regimens. Compositions of theinvention may also provide a faster rate of onset of exogenous SAMe incomparison to conventional oral dosage forms potentially leading toimprovements in efficacy.

BACKGROUND OF THE INVENTION

S-adenosyl-L-methionine (“SAMe”) is a naturally occurring compound thatis present in tissues throughout the body. At the molecular level, SAMeis involved in various metabolic pathways, including transmethylation,transsulfuration and aminopropylation.

In the body, SAMe is synthesized from an amino acid, methionine, and atriphosphate nucleotide, ATP. SAMe has been tested in numerous clinicaltrials for the treatment of various ailments, including arthritis, liverdisease and depression.

SAMe supplementation was initially considered impractical, due to theinstability of the SAMe ion during manufacturing, shipping and storage.Eventually stable salts of SAMe were developed (such as SAMe tosylatedisulfate, the butanedisulfonate salt of SAMe, the di-para-toluenesulfonate disulfate of SAMe, the tri-para-toluene sulfonic acid salt ofSAMe and the like). These salts can be formulated using standard, knowntechnologies used for non-parenteral administration including but notlimited to tablets, capsules and pellets. Formulations such as these mayalso comprise a coating which can serve multiple purposes such asreducing stomach irritation, improving taste and ease of swallowing, aswell as stabilizing the encapsulated SAMe from elements such asmoisture. Stable salts of SAMe are described in, for example, U.S. Pat.Nos. 3,954,726 and 4,057,686, both of which are incorporated herein byreference in their entirety. Conventional SAMe API is supplied as amolecular entity comprising an ion along with several counter-ions. Forexample, SAMe ion plus a tosylate and 2 sulfonic acid counter-ions makeup commercially available adenosylmethioninedisulfate-p-toluenesulfonate (also referred to as SAMe tosylatedisulfate). When referring to SAMe dosing, it is currently accepted inthe art that the numerical dose (usually in milligrams) refers to theamount of SAMe ion which is administered. For example, reference to a“400 mg SAMe tablet” of the SAMe tosylate disulfate would include the400 mg of SAMe ion, another 370 mg of the counter-ions, and 200-300 mgof additional excipient to make up a final tablet weight of 1.0-1.1grams. Thus, for example, a 1600 mg oral dose of SAMe which is generallyreported in the art would typically be a dose of four such 1.0-1.1 gramtablets taken at one time. Alternatively, the same 1600 mg dose of SAMeion may also be accomplished by administration of other combinations ofmultiple tablets such as, sixteen 100 mg or eight 200 mg tablets of SAMeion taken at a given time. Conventional oral dosage forms of SAMe aremost commonly produced with about 400 mg of SAMe ion; above that, thelarger dosage form becomes difficult for swallowing considering thateven at 400 mg of SAMe ion the tablets are quite large at 1.0-1.1 grams.

Exogenous SAMe exposure may be measured by looking at multiplepharmacokinetic parameters, the most common being the C_(max), T_(max)and AUC. After non-parenteral administration of SAMe, its concentrationin the blood increases until it reaches a peak concentration, thismeasured in plasma is the C_(max), and the time taken to reach theC_(max) is termed, T_(max). The area under the (plasma concentration)curve, or AUC, is another useful measurement and represents the drugexposure in the systemic circulation over a period of time.

A few studies examining these pharmacokinetic parameters in humans havebeen recorded for SAMe. The role of intravenous (IV) versus oraladministration of SAMe has been investigated to a small extent as wellas the effect of repeat dosing over time. Giulidori et al., reportplasma drug levels and half-lives of SAMe after a single, IVadministered dose (Giulidori, P. et al., (1984) Eur. J. Clin. Pharmacol.27:119.) Another group looked at SAMe plasma levels after a single,orally administered dose (Stramentinoli, G. (1987) Am. J. Med. 83:35.) Arecent study examines SAMe pharmacokinetic parameters after one-day andfive-day doses of orally and IV administered SAMe tosylate disulfate(Yang, J. et al (2009) Clin. Therapeutics, 31 (2): 311.) The prior artindicates that the half-life of oral SAMe is short and that AUC valuesof oral formulations are low.

There exists a need in the art to generate non-parenteral SAMeformulations with improved pharmacokinetic profiles compared toconventional prior art SAMe dosage forms. For example, those which haveincreased C_(max) and/or AUC values as well as those which are morepotent and exhibit similar C_(max) and AUC values at low doses of SAMe.High C_(max) or AUC formulations may produce an increased biologicalresponse to SAMe supplementation and ‘high potency’ formulations wouldhave the benefit of a lower pill count and potentially increasedtolerability for desired C_(max) and/or AUC values.

SUMMARY OF THE INVENTION

The present inventors have discovered that the pharmacokinetic (PK)profile of exogenous SAMe can be significantly improved by designingdosage forms to release substantial amounts of SAMe within a particular“window” of dissolution. Formulations that release the vast majority ofSAMe extremely early (i.e. those exhibiting an initial “burst” of drug)and those that are slower in their drug release are unable to achieveimproved in vivo PK profiles of SAMe. The investigators here identifycompositions and methods that are designed to release SAMe within thisunexpected “window” of preferred drug release levels. Thus, in someexemplified embodiments, compositions that exhibit improved SAMe PKprofiles have targeted amounts of drug release within a defineddissolution “window”—this in vitro correlates to a specified timeinterval for preferred drug release and in vivo relates to transitionthrough a specific region of the gastrointestinal tract.

In some embodiments of the invention improved in vivo PK profiles aregenerated when combining exogenous SAMe with suitable excipients and/orprocessing parameters that impart specific product characteristics suchas, for example, thickness, water content, friability, hardness,disintegration or dissolution properties. Accordingly, exemplifiedembodiments of the present invention relate to non-parenteralcompositions and methods which exhibit improved pharmacokineticprofiles, specifically high in vivo SAMe C_(max) values and/or increasedAUC values, in comparison to conventional prior art SAMe dosage forms.In some exemplified embodiments, provided are improved PK SAMecompositions which exhibit a targeted amount of drug release over adesired range of locations within the gastrointestinal tract of a fastedindividual. In certain embodiments, targeted drug release is achieved byuse of one or more functional coatings such that the functional coatingallows for extensive dissolution of the composition at the precise timeinterval in vitro. In some embodiments, targeted drug releaseformulations are identified in vitro using low pH dissolution profiles.Low pH dissolution studies are performed at below the standard of pH6.8. Accordingly, the invention also provides an in vitro screeningmethod which utilizes specific dissolution profiles of formulationcandidates to identify products which yield improved pharmacokineticvalues in vivo. Standard dissolution methods do not effectivelydistinguish these improved PK formulations from others. Obtainingdissolution profiles at low pH values (mimicking the pH of a specificlocation within the duodenum or upper small intestine where theformulations of the invention are targeted to release) in comparison todissolution profiles at pH 6.8 (which best represents the pH of thedistal small intestine) identifies rapid, yet targeted, dissolutionformulations as leading to improved pharmacokinetic parameters in vivo.

In other exemplified embodiments, compositions which exhibit improvedSAMe PK profiles are generated under conditions of very low relativehumidity. It is generally known that SAMe should be manufactured underconditions of low humidity (less than about 35%) in order to makeproducts of workable consistency. However, the investigators here havefound that SAMe formulations generated when humidity is maintained atvery low conditions, below about 10%, exhibit additional benefits suchas improved PK profiles which are less affected by variations inadditional parameters such as coating thickness.

There is provided herein a composition comprising a physiologicallyeffective dosage of SAMe, wherein non-parenteral administration of saidcomposition to a selected subject group produces in said selectedsubject group an effect comprising one of: a. an average maximum SAMeblood plasma concentration (average C_(max)) of at least about 1800ng/mL for a 1600 mg dosage of SAMe ion; b. an average SAMe plasma areaunder the curve (average AUC) of at least about 7500 ng·h/mL for a 1600mg dosage of SAMe ion; or, c. an average maximum SAMe blood plasmaconcentration (average C_(max)) of at least about 850 ng/mL and/or anaverage SAMe plasma area under the curve (average AUC) of at least about4000 ng·h/mL for a 800 mg dosage of SAMe ion; or, d. an average maximumSAMe blood plasma concentration (average Cmax) of at least about 400ng/mL and/or an average SAMe plasma area under curve (average AUC) of atleast about 1800 ng·h/mL for a 400 mg dosage of SAMe ion; or, e. anaverage maximum SAMe blood plasma concentration (average Cmax) of atleast about 200 ng/mL and/or an average SAMe plasma area under curve(average AUC) of at least about 900 ng·h/mL for a 200 mg dosage of SAMeion; or, f. an average maximum SAMe blood plasma concentration (averageCmax) of at least about 100 ng/mL and/or an average SAMe plasma areaunder curve (average AUC) of at least about 450 ng·h/mL for a 100 mgdosage of SAMe ion. In some embodiments, the composition comprisesphysical or chemical dosage form characteristics which modulate one ofsaid average SAMe C_(max) and said average SAMe AUC. In someembodiments, the composition is in a dosage form manufactured at arelative humidity of less than 10%. In some embodiments, the compositionis in a dosage form that comprises a functional coating whichconstitutes about 5% or less of the total weight of the dosage form. Insome embodiments, the composition is in a dosage form that comprises afunctional coating and the functional coating constitutes from 1 to 5%of the total weight of the dosage form. In some embodiments, thefunctional coating is comprised of one or more separate coatings orlayers. In some embodiments, the one or more separate coatings or layersmay be an enteric coating, a time-release coating, a pH-dependentcoating or other as well as combinations of these. In some embodiments,the dosage form characteristics comprise one of hardness, thickness,friability, speed of disintegration, speed of dissolution, shape, size,density, coating and combinations thereof. In some embodiments, thedosage form characteristics are modulated by controllably manipulatingduring production or manufacturing of said composition one of physicalmixing specifications, drying time, pressing conditions, environmentalparameters and combinations thereof. In some embodiments, the dosageform characteristics comprise a targeted dissolution profile at pH 6.0.In some embodiments, the dosage is divided into two, three, four, ormore dosage units. In some embodiments, the selected subject group is agroup of selected human subjects. In some embodiments, the compositionwhen administered to a select subject group provides in said selectedsubject group an improved pharmacokinetic profile through: a reducedvariation of T_(max) and equivalent AUC to bi-daily dosing and/orreduced side effects through once a day dosing. In some embodiments, thecomposition when administered to a selected subject group provides insaid selected subject group an average C_(max) within the range of about100 ng/mL to about 500 ng/mL per 100 mg of SAMe ion, or within a rangeof 110 ng/mL to about 500 ng/mL per 100 mg of SAMe ion; or within arange of 120 ng/mL to about 500 ng/mL per 100 mg of SAMe ion. In someembodiments, the composition when administered to a selected subjectgroup provides in said selected subject group an average AUC within therange of about 450 ng·h/mL to about 800 ng·h/mL for a 100 mg dosage ofSAMe ion. In some embodiments, the composition when administered to asubject provides in the subject one of an average T_(max) or C_(max)with reduced variation or a reduced effective dose in comparison to aSAMe reference data set. In some embodiments, the composition comprisesan oral delivery system, or a transmucosal delivery system. In someembodiments, the composition comprises one of tablets, pastes, capsules,granules, caplets, lozenges, pastes, and suppositories. In someembodiments, the composition comprises an oral delivery system. In someembodiments, dissolution of the oral delivery system or dosage formprovides about 20-90% release of SAMe after 60 minutes of being in anaqueous buffer having an initial pH of about 6. In some embodiments,dissolution of the oral delivery system or dosage form provides about25-80% release of SAMe after 60 minutes of being in an aqueous bufferhaving an initial pH of about 6. In some embodiments, dissolution of theoral delivery system or dosage in a USP II dissolution apparatus inaqueous buffer having initial pH of about 6.0 provides about 30-70%release of SAMe after 60 minutes of being in the buffer phase. Accordingto USP standards for dissolution profiling of an enteric-coated dosageform, a two hour incubation in an acidic/fluid phase precedes incubationin the aqueous buffer. Thus in some embodiments dissolution of the oraldelivery system or dosage form provides about 20-90% release of SAMeafter 60 minutes of being in an aqueous buffer having an initial pH ofabout 6, wherein prior to incubation in said aqueous buffer the oraldelivery system or dosage form is incubated for two hours in an acidicsolution. In some embodiments dissolution of the oral delivery system ordosage form provides about 25-80% release of SAMe after 60 minutes ofbeing in an aqueous buffer having an initial pH of about 6, whereinprior to incubation in said aqueous buffer the oral delivery system ordosage form is incubated for two hours in an acidic solution. In someembodiments, dissolution of the oral delivery system or dosage in a USPII dissolution apparatus in aqueous buffer having initial pH of about6.0 provides about 30-70% release of SAMe after 60 minutes of being inthe buffer phase, wherein prior to incubation in said buffer phase theoral delivery system or dosage form is incubated for two hours in anacidic solution. In some embodiments, the composition comprises adietary supplement. In some embodiments, the composition comprises amedical food. In some embodiments, there is provided a method oftreating a disease condition or disorder, comprising administering to asubject in need of such treatment an effective amount of the compositionof described herein. In some embodiments, the subject is human.

There is also provided herein an oral dosage composition comprising aphysiologically effective dosage of SAMe in combination with at leastone excipient, wherein administration of said composition to a selectedsubject group produces in said selected subject group an effectcomprising one of: a. an average maximum SAMe blood plasma concentration(average C_(max)) of at least about 1800 ng/mL for a 1600 mg dosage ofSAMe ion; or, b. an average SAMe plasma area under the curve (averageAUC) of at least about 7500 ng·h/mL for a 1600 mg dosage of SAMe ion;or, c. an average maximum SAMe blood plasma concentration (averageC_(max)) of at least about 850 ng/mL and an average SAMe plasma areaunder the curve (average AUC) of at least about 4000 ng·h/mL for a 800mg dosage of SAMe ion; or, d. an average maximum SAMe blood plasmaconcentration (average C_(max)) of at least about 400 ng/mL and/or anaverage SAMe plasma area under curve (average AUC) of at least about1800 ng·h/mL for a 400 mg dosage of SAMe ion; or, e. an average maximumSAMe blood plasma concentration (average C_(max)) of at least about 200ng/mL and/or an average SAMe plasma area under curve (average AUC) of atleast about 900 ng·h/mL for a 200 mg dosage of SAMe ion; or, f. anaverage maximum SAMe blood plasma concentration (average C_(max)) of atleast about 100 ng/mL and/or an average SAMe plasma area under curve(average AUC) of at least about 450 ng·h/mL for a 100 mg dosage of SAMeion. In some embodiments, the at least one excipient is one of matrixmaterials; binders; lubricants, glidants, coatings, disintegrants,super-disintegrants, polysaccharides, oligosaccharides, polypeptides,proteins, synthetic oligomers, synthetic polymers, monomeric organicmolecules, hydrophobic organic molecules, hydrophilic organic molecules,amphoteric organic molecules, inorganic salts, inorganic metals, andcombinations thereof. In some embodiments, the composition comprisesphysical or chemical dosage form characteristics which modulate one ofsaid average SAMe C_(max) and said average SAMe AUC. In someembodiments, the composition is in a dosage form manufactured at arelative humidity of less than 10%. In some embodiments, the compositionis in a dosage form that comprises a functional coating and thefunctional coating constitutes 5% or less of the total weight of thedosage form. In some embodiments, the composition is in a dosage formthat comprises a functional coating and the functional coatingconstitutes from 1 to 5% of the total weight of the dosage form. In someembodiments, the functional coating is comprised of one or more separatecoatings or layers. In some embodiments, the one or more separatecoatings or layers are each an enteric coating, a time-release coating,a pH-dependent coating or other as well as combinations of these. Insome embodiments, the dosage form characteristics comprises one ofhardness, thickness, friability, speed of disintegration, speed ofdissolution, shape, size, density, coating and combinations thereof. Insome embodiments, the dosage form characteristics are modulated bycontrollably manipulating during production of said composition one ofphysical mixing specifications, drying time, pressing conditions,environmental parameters, and combinations thereof. In some embodiments,the composition is manufactured under specific conditions comprising oneof mixing method (including sieve size, rpm, and milling), drying time,press conditions, environmental parameters and combinations thereof. Insome embodiments, the dosage is divided into two, three, four, or moredosage units. In some embodiments, the selected subject group is aselected group of humans. In some embodiments, the composition whenadministered to a selected subject group provides in said selectedsubject group an average C_(max) within the range of about 100 ng/mL toabout 500 ng/mL per 100 mg of SAMe ion. In some embodiments, thecomposition when administered to a selected subject group provides insaid selected subject group an average AUC within the range of about 450ng·h/mL to about 800 ng·h/mL for a 100 mg dosage of SAMe ion. In someembodiments, the composition when administered to a subject provides inthe subject one of an average T_(max) or C_(max) with reduced variationor a reduced effective dose in comparison to a SAMe reference data set.In some embodiments, there is further provided a method of treating adisease condition or disorder comprising administering to a subject inneed of such treatment an effective amount of the composition describedherein. In some embodiments, the subject is a human.

Also provided herein is a dietary supplement preparation comprising aphysiologically effective dosage of SAMe in combination with at leastone excipient, wherein administration of said composition to a selectedsubject group produces in said selected subject group an effectcomprising one of: a. an average maximum SAMe blood plasma concentration(average C_(max)) of at least about 1800 ng/mL for a 1600 mg dosage ofSAMe ion; or, b. an average SAMe plasma area under the curve (averageAUC) of at least about 7500 ng·h/mL for a 1600 mg dosage of SAMe ion;or, c. an average maximum SAMe blood plasma concentration (averageC_(max)) of at least about 850 ng/mL and an average SAMe plasma areaunder the curve (average AUC) of at least about 4000 ng·h/mL for a 800mg dosage of SAMe ion; or, d. an average maximum SAMe blood plasmaconcentration (average C_(max)) of at least about 400 ng/mL and/or anaverage SAMe plasma area under curve (average AUC) of at least about1800 ng·h/mL for a 400 mg dosage of SAMe ion; or, e. an average maximumSAMe blood plasma concentration (average C_(max)) of at least about 200ng/mL and/or an average SAMe plasma area under curve (average AUC) of atleast about 900 ng·h/mL for a 200 mg dosage of SAMe ion; or, f. anaverage maximum SAMe blood plasma concentration (average C_(max)) of atleast about 100 ng/mL and/or an average SAMe plasma area under curve(average AUC) of at least about 450 ng·h/mL for a 100 mg dosage of SAMeion. In some embodiments, the at least one excipient is one of matrixmaterials; binders; lubricants; glidants; coatings; disintegrants,super-disintegrants; polysaccharides, oligosaccharides; polypeptides,proteins synthetic oligomers, synthetic polymers, monomeric organicmolecules, hydrophobic organic molecules, hydrophilic organic molecules,amphoteric organic molecules, inorganic salts inorganic metals, andcombinations thereof. In some embodiments, the composition comprisesphysical or chemical dosage form characteristics which modulate one ofsaid average SAMe C_(max) and said average SAMe AUC. In someembodiments, the composition is in a dosage form manufactured at arelative humidity of less than 10%. In some embodiments, the compositionis in a dosage form that comprises a functional coating and thefunctional coating constitutes 5% or less of the total weight of thedosage form. In some embodiments, the composition is in a dosage formthat comprises a functional coating and the functional coatingconstitutes from 1 to 5% of the total weight of the dosage form. In someembodiments, the functional coating is comprised of one or more separatecoatings or layers. In some embodiments, the one or more separatecoatings or layers are each an enteric coating, a time-release coating,a pH-dependent coating or other as well as combinations of these. Insome embodiments, the dosage form characteristics comprises one ofhardness, thickness, friability, speed of disintegration, speed ofdissolution, shape, size, density, coating and combinations thereof. Insome embodiments, the dosage form characteristics are modulated bycontrollably manipulating during production of said composition one ofphysical mixing specifications, drying time, pressing conditions,environmental parameters and combinations thereof. In some embodiments,the composition is manufactured under specific conditions comprising oneof mixing method (including sieve size, rpm, and milling), drying time,press conditions, environmental parameters and combinations thereof. Insome embodiments, the selected subject group is a selected group ofhumans. In some embodiments, the composition when administered to aselected subject group provides in said selected subject group anaverage C_(max) within the range of about 100 ng/mL to about 500 ng/mLper 100 mg of SAMe ion. In some embodiments, the composition whenadministered to a selected subject group provides in said selectedsubject group an average AUC within the range of about 450 ng·h/mL toabout 800 ng·h/mL for a 100 mg dosage of SAMe ion. In some embodiments,the composition when administered to a subject provides in the subjectone of an average T_(max) or C_(max) with reduced variation or a reducedeffective dose in comparison to a SAMe reference data set. In someembodiments, there is provided a method of treating a disease conditionor disorder comprising administering to a subject in need of suchtreatment an effective amount of a composition described herein. In someembodiments, the subject is a human.

Also provided herein is a method for improving the pharmacokineticparameters of exogenous SAMe administered to a subject, said methodcomprising administering to the subject a non-parental compositioncomprising at least one physiologically effective dosage of SAMe incombination with at least one excipient selected to improve thepharmacokinetic parameters of said SAMe in a subject, saidpharmacokinetic parameters measurable in the subject by one of aC_(max), an AUC, and combinations thereof in comparison to a selectedSAMe reference data set. In some embodiments, the at least one excipientis one of matrix materials; binders; lubricants; glidants; coatings;disintegrants, super-disintegrants; polysaccharides, oligosaccharides;polypeptides, proteins synthetic oligomers, synthetic polymers,monomeric organic molecules, hydrophobic organic molecules, hydrophilicorganic molecules, amphoteric organic molecules, inorganic saltsinorganic metals, and combinations thereof. In some embodiments, theimprovement of the pharmacokinetic parameters of said SAMe is a functionof a physical or chemical dosage form characteristic of the composition.In some embodiments, the composition is manufactured at a relativehumidity of less than 10%. In some embodiments, the composition is in adosage form, which includes a functional coating, and the functionalcoating accounts for 5% or less of the total weight of the dosage form.In some embodiments, the functional coating accounts for 1% to 5% of thetotal weight of the dosage form. In some embodiments, the physical orchemical dosage form characteristic comprises one of hardness,thickness, friability, speed of disintegration, speed of dissolution,shape, size, coating, density, and combinations thereof. In someembodiments, the composition when administered to a selected subjectgroup provides in the selected subject group an average C_(max) of atleast about 1800 ng/mL for a 1600 mg dosage of SAMe ion. In someembodiments, the composition when administered to a selected subjectgroup provides in the selected subject group an average C_(max) of atleast about 850 ng/mL and an average SAMe plasma area under the curve(average AUC) of at least about 4000 ng·h/mL for a 800 mg dosage of SAMeion. In some embodiments, the composition when administered to aselected subject group provides in the selected subject group an averageC_(max) of at least about 100 ng/mL per 100 mg of SAMe ion in saidphysiologically effective dosage. In some embodiments, the compositionwhen administered to a selected subject group provides in the selectedsubject group an average C_(max) within the range of about 110 ng/mL toabout 500 ng/mL per 100 mg of SAMe ion in said physiologically effectivedosage. In some embodiments, the composition when administered to aselected subject group provides in the selected subject group one of anaverage T_(max) with reduced variation or a reduced effective dose incomparison to a SAMe reference data set. In some embodiments, thecomposition when administered to a selected subject group provides inthe selected subject group an average AUC of at least about 7500 ng·h/mLfor a 1600 mg dosage of SAMe ion. In some embodiments, the compositionwhen administered to a selected subject group provides in the selectedsubject group an average AUC of at least about 4000 ng·h/mL for a 800 mgdosage of SAMe ion. In some embodiments, the composition whenadministered to a selected subject group provides in the selectedsubject group an average AUC within the range of about 500 ng·h/mL toabout 800 ng·h/mL for a 100 mg dosage of SAMe ion. In some embodiments,the subjects comprising the selected subject group are one of humans,livestock animals, exotic animals, avian species, laboratory animals,canines, felines, and primates.

In some embodiments, there is provided a method of treating in a patienta disease or disorder selected from the group consisting of mental andpsychiatric disorders, nervous system diseases and disorders,neurological diseases and disorders, conditions associated with injuriesto the central nervous system, liver diseases and disorders, cancers,joint diseases and disorders, inflammatory diseases and disorders,autoimmune diseases and disorders, degenerative diseases and disorders,soft-tissue diseases and disorders, pain diseases and disorders,cardiovascular disorders related to hyper-homocysteinemia andhypo-homocysteinemia, genetic disorders related to hyper-methylation andhypo-methylation, gastrointestinal diseases and disorders, and disordersinduced in whole or in part by oxidative or free-radical damage,comprising administering to the patient in need thereof a composition asdescribed herein.

There is also provided herein a formulation comprising SAMe, wherein theformulation comprises a mixture of SAMe and at least one excipient andthe mixture is produced by combining said SAMe and said excipient at arelative humidity less than about 10%.

There is also provided herein a formulation comprising SAMe, wherein theformulation comprises a mixture of SAMe and at least one excipient,wherein the mixture exhibits a dissolution profile at pH 6.0 suitable totarget delivery to the proximal intestine.

There is also provided herein a process of improving the pharmacokineticprofile of a SAMe formulation, comprising manufacturing said SAMeformulation at a relative humidity of less than about 10%.

There is also provided herein a composition for oral administration,comprising SAMe and at least one excipient wherein the formulationexhibits an in vitro dissolution profile in pH 6.0 aqueous solution suchthat greater than 20% and less than 90% of total SAMe in the compositionis dissolved from 30 to 90 minutes of incubation in said pH 6.0 aqueoussolution. In some embodiments, the formulation exhibits an in vitrodissolution profile in pH 6.0 aqueous solution such that greater than25% and less than 80% of total SAMe in the composition is dissolved from45 to 75 minutes of incubation in said pH 6.0 aqueous solution. Prior toincubation in said pH 6.0 aqueous solution the formulation is incubatedfor about 2 hours in an acidic phase as according to USP standards fordissolution testing of enteric-coated dosage forms. In some embodiments,the composition is in a dosage form manufactured at a relative humidityof less than 10%. In some embodiments, the composition is in a dosageform that comprises a functional coating and the functional coatingconstitutes 5% or less of the total weight of the dosage form. In someembodiments, the composition is in a dosage form that comprises afunctional coating and the functional coating constitutes from 1 to 5%of the total weight of the dosage form. In some embodiments, thefunctional coating is comprised of one or more separate coatings orlayers that together constitute about 5% or less of the total weight ofthe dosage form.

There is also provided herein a method for improving the uptake of SAMe,comprising administering to a patient SAMe in a formulation thatexhibits an in vitro dissolution profile at pH 6.0, wherein greater than20% and less than 90% of total SAMe is dissolved between 30 to 90minutes of incubation in the pH 6.0 aqueous buffer. In some embodiments,the formulation exhibits an in vitro dissolution profile in pH 6.0aqueous solution such that greater than 25% and less than 80% of totalSAMe in the composition is dissolved from 45 to 75 minutes of incubationin the pH 6.0 aqueous buffer. In some embodiments, the composition is ina dosage form manufactured at a relative humidity of less than 10%. Insome embodiments, the composition is in a dosage form that comprises anenteric coating and the enteric coating constitutes 5% or less of thetotal weight of the dosage form. In some embodiments, the composition isin a dosage form that comprises an enteric coating and the entericcoating constitutes from 1 to 5% of the total weight of the dosage form.

For greater clarity all references to dose within this patent refer todose as the dose of SAMe ion. Pharmacokinetic parameters such as averagemaximum plasma concentration of SAMe (C_(max)) are determined using abioanalytical method with adequate sensitivity, specificity, ruggedness,stability and repeatability (for example, a qualified liquidchromatography triple quad mass spectrometry based method coupled with asuitable extraction method for the separation of analyte from plasma).AUC values were calculated from 0-24 hours using the trapezoid methodand are uncorrected for baseline, endogenous SAMe levels. A suitable“selected subject group” has six or more subjects who are dosed fasted.All members of the “selected subject group” have pharmacokineticparameters for SAMe that fall within statistically normal ranges (i.e.no outliers) and no member will be included on the basis of non-standardor unusual SAMe absorption or metabolism. The average C_(max) values arederived by averaging the concentration at each time point for allmembers of the subject group. Use of methods of the invention in vivoprovides high C_(max) and/or AUC values in comparison to conventionaldosage forms of SAMe.

Some embodiments of the invention also relate to compositions andmethods which yield a lower effective dose and/or less variablepharmacokinetic parameters (such as T_(max) values with reducedvariation) in comparison to conventional non-parenteral SAMeformulations. A “lower effective dose” or “reduced effective dose” ismeant to define a physiologically acceptable dose of exogenous SAMewhich results in pharmacokinetic parameters which are equivalent to asignificantly higher dose of another non-parenteral SAMe formulation,such as that obtained through administration of a higher dose of one ormore currently commercially available SAMe formulations. Formulationssuch as these which exhibit similar C_(max) and AUC values at lower SAMedoses would have many benefits including a lower pill burden andpotentially increased tolerability.

Additional embodiments of the invention also relate to compositions andmethods which yield an improved side effect profile in comparison toconventional non-parenteral SAMe formulations. An “improved side effect”or “reduced side effect” or “beneficial side effect” profile is meant todefine improved tolerability to administration of exogenous SAMe, suchas less frequency and/or reduced intensity of side effects associatedwith SAMe supplementation.

Some exemplary embodiments of the present invention also relate to adosing regimen of SAMe of once daily, or QD dosing, which results inimproved pharmacokinetic profiles while delivering similar or greaterAUC levels of SAMe to the subject in comparison to conventional twicedaily or more frequent dosing. In certain embodiments, the effect ofonce a day dosing is believed to result in the most consistentpharmacokinetic parameter measurements, specifically those of theC_(max) and T_(max). The less variable pharmacokinetic profiles thatresult from once a day dosing of these formulations allow for morecertainty of dosing and exposure by the medical practitioner as well asimproved side effect profiles for subjects.

In some embodiments of the present invention formulations which exhibitsuperior pharmacokinetic profiles in comparison to conventionalnon-parenteral SAMe dosage forms provide an improved rate of onset ofSAMe which may result in enhanced therapeutic outcomes.

Other exemplary embodiments of the invention relate to methods fortreating a disease or disorder in a subject and/or improving thenutritional status in a subject, said methods comprising administeringto said subject compositions of the invention comprising physiologicallyeffective dosages of SAMe thereby improving the pharmacokinetic profileof SAMe. Improved pharmacokinetic profiles are identified by, forexample, an increase in C_(max) and/or AUC values; or alternatively adecrease in effective dose; or pharmacokinetic parameters with reducedvariation. Achieving one or more of these criteria would constitute animprovement in the pharmacokinetic profile of SAMe.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph of the average SAMe plasma concentration with thestandard error of the mean and showing the average maximum plasmaconcentration (C_(max)) of subjects administered a single 1600 mg SAMeion dose from either a commercially available oral formulation ofS-adenosyl methionine tosylate disulphate (open squares) or a single1600 mg oral dose of MSI-43 of the present invention (closed circles);

FIG. 2A is a graph of the dissolution profiles of various SAMe tabletformulations of the invention at pH 6.8 represented as the percent drugreleased over time (hours); and,

FIG. 2B is a graph of the pH 6.0 dissolution profiles of the same SAMetablet formulations of the invention as seen in FIG. 2A represented asthe percent drug released over time (hours);

FIG. 3 is a graph of the pH 6.0 dissolution profiles of additional SAMeformulations of the invention in comparison to two commerciallyavailable SAMe tablets;

FIG. 4 is a graph showing the glass transition temperature (Tg) of SAMeas a function of relative humidity (RH).

FIG. 5A is a graph of the average SAMe plasma concentration with thestandard error of the mean and showing the average maximum plasmaconcentration (C_(max)) from seven subjects who were administered eithera 1600 mg (SAMe ion) oral dose of SAMe formulations of the invention(MSI-72; open squares) or a 1600 mg (SAMe ion) oral dose of acommercially available oral formulation of SAMe (closed squares); and,

FIG. 5B is a graph of the average SAMe plasma concentration with thestandard error of the mean and showing the maximum plasma concentration(C_(max)) from seven subjects who were administered either an 800 mg(SAMe ion) oral dose of the SAMe formulations of the invention reportedin FIG. 5A (open squares) or a 1600 mg (SAMe ion) oral dose of the samecommercially available oral formulation of SAMe also detailed in FIG. 5A(closed squares); and,

FIG. 5C is a graph of the average SAMe plasma concentration with thestandard error of the mean and showing the maximum plasma concentration(C_(max)) from seven subjects who were administered either an 800 mg(SAMe ion) oral dose of a different SAMe formulation of the invention,MSI-69 (open squares) or a 1600 mg (SAMe ion) oral dose of the samecommercially available oral formulation of SAMe also detailed in FIG. 5A(closed squares);

FIG. 6 is a graph of the average SAMe plasma concentration with thestandard error of the mean and showing the pharmacokinetics of subjectsgiven 800 mg (SAMe ion) of a commercially available oral formulation ofS-adenosyl methionine tosylate disulphate wherein the subjects wereeither fed (open circles) or fasted (closed squares) prior toadministration of the 800 mg dose;

FIG. 7 is a graph with two pharmacokinetic profiles, both of the averageSAMe plasma concentration. This graph, based on the data shown in FIG.6, is a simulation of an 800 mg BID (twice daily) dosing separated by 8hours. The subjects were dosed with 800 mg (SAMe ion) of a commerciallyavailable oral formulation of S-adenosylmethionine disulfate tosylateunder fasted (closed squares) or fed (open circles) conditions. 8 hourswas added to each of the time points for the fed dataset to simulate a4:00 pm dosing.

DETAILED DESCRIPTION OF THE INVENTION

The present inventors have discovered that the pharmacokinetic (PK)profile of exogenous SAMe in plasma can be significantly improved bydesigning dosage forms to release substantial amounts of drug within aparticular “window” along the path of dissolution. Surprisingly,formulations which release the vast majority of SAMe extremely early(i.e. those exhibiting an initial “burst” of drug) and those which areslower in their drug release are unable to achieve improved in vivo PKprofiles of SAMe. The investigators here identify compositions andmethods which are designed to release SAMe within this unexpected“window” of preferred drug release levels. Thus, in some exemplifiedembodiments, compositions which exhibit improved SAMe PK profiles havetargeted amounts of drug release within a particular dissolution“window”. This targeted dissolution is such that extensive drug releaseoccurs rapidly over a defined period of time as observed using low pHdissolution studies in vitro. Some exemplary embodiments of theinvention therefore also relate to an in vitro screening method ofidentifying formulations which exhibit improved PK profiles in vivo.Screening is carried out by performing dissolution studies at pH valueslower than the standard of pH 6.8.

In other embodiments, compositions which exhibit improved SAMe PKprofiles are generated under conditions of very low relative humidity.Additional embodiments of the invention relate to non-parenteralcompositions and methods that improve the PK profile of exogenous SAMeand methods of using the same, e.g. for the treatment of variousdiseases or disorders in a subject and/or improving the nutritionalstatus of a subject. In certain embodiments compositions of theinvention are administered to the subject once per day. In someembodiments, administration of compositions of the invention to asubject results in an improved side effect profile of said subject. Inother embodiments, compositions of the invention provide rapid onset ofexogenous SAMe in comparison to conventional non-parenteral SAMe dosageforms.

Further embodiments of the invention relate to combinations of SAMe withone or more active ingredients that are commonly prescribed or used fortreatment of and/or prophylaxis of various diseases or disorders in asubject. Some other embodiments of the invention relate tonon-parenteral compositions and methods that improve the PK profile ofexogenous SAMe either alone or in combination with one or moreadditional agents and additionally improve the side effect profileassociated with SAMe and/or said one or more additional agents.

“Improved pharmacokinetic profile” or “enhanced pharmacokinetic profile”as used herein refers to one or more of the following criteria incomparison to conventional oral SAMe treatments: 1) high average C_(max)(greater than about 1800 ng/mL when tested at a dose of 1600 mg (SAMeion)); and/or 2) increased AUC (greater than about 7500 ng·h/mL whentested at a dose of 1600 mg (SAMe ion) or greater than about 4000ng·h/mL when tested at a dose of 800 mg); and/or 3) pharmacokineticparameters with reduced variation; and/or 4) reduced effective dose (forexample, a C_(max) of at least about 100 ng/mL per 100 mg dose of SAMeion and/or an AUC of about 450 ng·h/mL per 100 mg dose of SAMe ion). Insome embodiments, the invention provides a blood plasma SAMe C_(max) ofgreater than about 2000 ng/mL, greater than about 3000 ng/mL, or greaterthan about 3500 ng/mL when tested at a dose of 1600 mg (SAMe ion). Insome embodiments, the invention provides a C_(max) of at least about 110ng/mL per 100 mg dose of SAMe ion, at least about 130 ng/mL per 100 mgdose of SAMe ion, at least about 150 ng/mL per 100 mg dose of SAMe ion,at least about 180 ng/mL per 100 mg dose of SAMe ion, at least about 210ng/mL per 100 mg dose of SAMe ion, or at least about 240 ng/mL per 100mg of SAMe ion. In some embodiments, the invention provides an AUC ofgreater than about 8000 ng·h/mL, greater than about 10000 ng·h/mL,greater than about 11000 ng·h/mL, or greater than about 12000 ng·h/mLwhen tested at a dose of 1600 mg (SAMe ion). In some embodiments, theinvention provides an AUC of at least 500 ng·h/mL per 100 mg SAMe dosed,at least 600 ng·h/mL per 100 mg SAMe dosed, at least 700 ng·h/mL per 100mg SAMe dosed, or at least 800 ng·h/mL per 100 mg SAMe dosed.

As used herein the term “SAMe” refers to S-adenosyl-L-methionine (or,more simply, “S-adenosylmethionine”). When referring to dose, the amount(typically in mg) refers to the dose of SAMe ion administered. As shownin the structural formula presented earlier, SAMe appears as a chargedspecies, and its ionization state varies with pH. In its solid form,SAMe is most commonly available as a stable salt form, e.g. withp-toluenesulfonic acid alone or in combination with one or moreadditional salt-forming substances for example, mineral or organic acidsand/or amino acids. (See U.S. Pat. No. 3,893,999, incorporated herein byreference in its entirety). Other stable SAMe salts are described in,for example, U.S. Pat. No. 5,128,249, which describes particular stablesalts of SAMe. Various morphologies of SAMe are suitable for use in thepresent invention. Thus, as used herein “SAMe” refers to the stablesalts, amorphous forms, semicrystalline forms and crystalline forms ofSAMe as well as to the ionic form of SAMe when present in vivo. A“physiologically effective dosage” of SAMe as used herein is meant toinclude an amount of SAMe which is administered under a defined dosingregimen for either clinical, pharmaceutical, medicinal, veterinary,dietary or nutritional purposes. Thus a “physiologically effectivedosage” or a “physiologically acceptable dosage” of SAMe includes atherapeutically effective dosage, a pharmaceutically acceptable dosage,a veterinary acceptable dosage, a nutraceutically acceptable dosage, adietary acceptable dosage and a nutritionally acceptable dosage of SAMeas well as an acceptable dosage for use as a medical food and all ofwhich are included for use in the present invention. When referring to“medicinal” preparations, purposes or treatments they are meant toinclude “medical foods”. Medical foods are defined by the U.S. Food andDrug Administration as a food which is formulated to be consumed oradministered enterally under the supervision of a physician and which isintended for the specific dietary management of a disease or conditionfor which distinctive nutritional requirements, based on recognizedscientific principles, are established by medical evaluation.

Some exemplary embodiments of the present invention relate tonon-parenteral compositions and methods of their use for enhancing theeffectiveness of a physiologically effective dosage of SAMe utilized asa medical food or dietary or nutritional supplement in a subject.

Some exemplary embodiments of the invention relate to a method fortreating and/or prophylaxis in a subject a disease or disorder selectedfrom the group consisting of, but not limited to, a mental orpsychiatric disorder (e.g. psychotic/mood or non-psychotic mentaldisorders exemplified by depression and substance related disorders,respectively), a nervous system disease/disorder (e.g. a central nervoussystem disease exemplified by Alzheimer's), other neurologicaldisease/disorders (e.g. headaches and sleep disorders), conditionsassociated with injury to the central nervous system, a liverdisease/disorder (e.g. alcoholic liver disease), a cancer (e.g. solidand blood-borne cancers), a joint disease/disorder (e.g. arthritis), aninflammatory disease/disorder (e.g. ulcerative colitis), an autoimmunedisease/disorder (e.g. systemic lupus erythematosis and rheumatoidarthritis), a degenerative disease/disorder (e.g. Amyotrophic LateralSclerosis), a soft-tissue disease/disorder (e.g. a fibromyalgiadisorder), a pain disease/disorder, a genetic disorder related to hyper-or hypo-methylation, a gastrointestinal disease/disorder, acardiovascular disease/disorder, and a disorder induced in whole or inpart by oxidative or free-radical damage, comprising administering tosaid subject an exemplary composition of the present invention whichprovides a physiologically effective amount of exogenous SAMe with animproved PK profile.

Some exemplary embodiments of the present invention relate tonon-parenteral compositions and methods which improve the side effectprofile associated with a physiologically effective amount of exogenousSAMe.

Other exemplary embodiments of the present invention relate tocombinations of SAMe with one or more active ingredients that arecommonly prescribed or used for treating and/or prophylaxis in a subjecta disease or disorder selected from the group consisting of, but notlimited to, those detailed above. In some embodiments administration ofimproved PK SAMe formulations of the invention with one or more activeingredients that are commonly prescribed or used for treating and/orprophylaxis in a subject a disease or disorder selected from the groupdescribed above leads to an improved side effect profile of the subject.In certain embodiments, the side effect profile resulting from use ofsaid one or more active ingredients that are commonly prescribed or usedfor treating and/or prophylaxis in a subject a disease or disorderselected from the group described above, is improved.

SAMe Formulations for Non-Parenteral Administration

Formulations for non-parenteral administration of drugs/therapeuticagents are typically provided as solid or semi-solid products or dosageforms, exemplified by tablets, capsules or pellets, and generallyconsist of a core “matrix material” which ‘encapsulates’ the drug aswell as one or more protective coatings. “Product” or “dosage form” asused herein refers to any solid or semi-solid formulation or preparationused for non-parental administration. Non-parenteral formulations orpreparations as described herein include oral delivery systemsexemplified by tablets, pastes, capsules, granules, caplets, lozengesand the like; and transmucosal or inhaled delivery systems, exemplifiedby aerosols, irrigants, topical creams, pastes, lozenges, patches, andthe like, all of which are well-known and well-documented in the art.These formulations may be administered using a clinical, pharmaceuticalor veterinary dosing regimen. Non-parenteral dosage forms may also beprovided as medical foods or dietary or nutritional supplements.Non-parenterally administered SAMe formulations may be configured toenable extended release of the formulated SAMe. Co-owned U.S. patentapplication 2009/0088404, which is incorporated herein by reference,provides novel formulations of extended-release SAMe formulations.

Upon administration, the rate of release of an active moiety from anon-parenteral product can be greatly influenced by the excipientsand/or product characteristics which make up the product itself. Forexample, an enteric coat on a tablet is designed to separate thattablet's contents from the stomach contents to prevent, for example,degradation of the stomach which may induce gastrointestinal discomfortor injury. SAMe and other tablets described in the art are commonlyenteric coated. Once the dosage form has transited from the stomach tothe duodenum and subsequently the rest of the small intestine theenteric coat is removed due the pH change and the tabledisintegrates/dissolves according to its intrinsic properties and thedosage form technologies that have been applied. According to thecurrently accepted conventional understanding, systemic exposure of theactive moiety will be relatively insensitive to the small formulationchanges. For example, following the conventional understanding, onewould expect similar or near similar PK behavior for formulations withan application of enteric coat within the normal range of recommendedapplication thicknesses. Similarly, following the conventionalunderstanding, one would expect comparable behavior for dosage formsprepared within a range of humidity that allowed for the efficient andeffective handling of the active moiety through the course of theformulation process. For SAMe as the active moiety, the impact of theseprocessing parameters is expected to be particularly blunted in light ofthe teaching within the art that SAMe is subject to low bioavailabilitydue to extensive first pass metabolism. Thus one would predictconsistent systemic exposure as measured by pharmacokinetics fordifferent enteric coated dosage forms within the normal range ofoperating parameters as long as active moiety was delivered intact tothe site of absorption in the small intestine. However, theinvestigators here have found that, contrary to conventionalunderstanding, combinations of exogenous SAMe with one or moreexcipients and/or processing parameters which result in specific productcharacteristics dramatically affect the pharmacokinetic profile of SAMeand lead to high C_(max) and AUC values in vivo.

Suitable excipients which result in improved pharmacokinetic profiles ofSAMe are preferably included in non-parenteral formulations of theinvention. More specifically, formulations which include SAMe and one ormore suitable excipients, exemplified by matrix materials, binders,lubricants, glidants or disintegrants which aid in modulating the PKprofile of administered exogenous SAMe are preferred. Other embodimentsof the invention relate to compositions comprising SAMe in combinationwith one or more suitable excipients and one or more specific productcharacteristics (such as dissolution or water content) which result inimproved pharmacokinetic profiles of SAMe in vivo. Thus, the in vivoperformance of non-parenteral SAMe dosage forms/products included hereinis based upon the composition of the excipients added duringmanufacturing and/or the final product characteristics generated throughspecific processing parameters and methods.

Product or Dosage Form Characteristics

The product or dosage form characteristics which result from theprocessing methods and/or parameters for generating non-parenteralformulations such as tablets, include, but are not limited to, hardness,thickness, water content, friability, disintegration, dissolutionprofile(s), shape, size, weight, uniformity and composition. Theseproduct characteristics can be modulated in a number of ways and affectthe final in vitro and/or in vivo performance of the formulations. As anexample, tablets generated by compression or molding processes may havevarying degrees of thickness or hardness depending on the processingparameters under which they were made. Product or dosage formcharacteristics may be a consequence of excipient selection, excipientcomposition, manufacturing methods applied or a combination of any ofthese. The combination of excipients as well as product characteristics(including processing methods or processing parameters) of the finaldosage form will ultimately determine the pharmacokinetic profile of theactive ingredient in vivo. The non-parenterally administered SAMeformulations of the invention may be processed or manufactured underspecific conditions such as, for example, mixing methods (includingsieve size, rpm, and milling), drying time, press conditions,environmental parameters (e.g. temperature and humidity) andcombinations thereof) which themselves modulate the pharmacokineticprofile of SAMe in vivo (i.e. increase the average C_(max) or AUC). Inorder to quantitatively compare one tablet to another, it is customaryto measure several of these product or dosage form characteristics. Thisis also necessary when attempting to duplicate multiple batches.

Surprisingly, the present investigators found that a specific “window”of dissolution (i.e. a particular amount of drug release over a certaintime frame) correlated with those formulations of the invention whichexhibited an improved SAMe PK profile in vivo. Although dissolutionstudies are commonly utilized to characterize non-parenteralformulations, testing is standard using a buffer phase which is at pH6.8 to best represent the pH of the distal small intestine. In addition,dissolution profiles are commonly referred to in the art as either“fast” or “slow”; however, the present investigators have identifiedthat in fact a specific “window” of fast dissolution leads to levels ofSAMe in the plasma not previously reported in these arts.

Dissolution and drug release from formulations depends on many factorsincluding the solubility and concentration of the active ingredient, thenature and composition of the excipients, content uniformity, watercontent, product shape and size, porosity, disintegration time and otherfactors. The release of a drug or active ingredient from a final dosageform in vitro is typically characterized by its dissolution profileunder standardized conditions (using United States Pharmacopeia (USP) orsimilar accepted methods for reference) and at pH 6.8 as mentionedabove. The dissolution profile shows the amount of drug released overtime into the test media under specified conditions. Standard conditionsmake use of buffers at pH 6.8 in order to best mimic the pH of distalsmall intestine. The dissolution test method for enteric dosage formsinvolves incubation of the formulation in a first acidic phase for twohours and is then transferred to the aqueous buffer phase (pH 6.8). Timepoints for measuring drug release begin at this two-hour time period(i.e. when first transferred into the aqueous buffer phase).Investigators here found that when dissolution profiles of multiple SAMeformulations of the invention are analyzed under conditions of pH 6.8,all formulations show “fast” dissolution and their dissolution profilescannot be distinguished from one formulation to the next. However, whenthe dissolution studies were conducted using a pH 6.0 buffer, which bestmimics that the pH of a specific region of the upper small intestinewhere SAMe is absorbed, there is significant differentiation between theformulations which are “fast” to dissolve and release drugs. Moreover,pharmacokinetic analysis of these formulations in vivo showed thatformulations of the invention dissolving rapidly within a specific“window”, as seen in the pH 6.0 dissolution profiles, correlated withthose formulations exhibiting very high C_(max) and AUC values (seeExamples 2-3).

Some embodiments of the invention thus relate to improvedpharmacokinetic SAMe compositions which show targeted dissolution in abuffer phase of pH 6.0. Preferably, between 25-80% SAMe is releasedafter one hour of incubation in the buffer phase; more preferably about30-70% SAMe is released within one hour of incubation in the bufferphase; and even more preferably, about 30-60% SAMe is released withinone hour of incubation in the buffer phase.

Excipients and Processing Parameters Suitable for Use in the Invention

Excipients are usually grouped by their function such as: disintegrants,diluents, binders, lubricants, glidants, coatings, coloring agents orflavoring agents, and the same excipient may be used for more than onefunction in a given oral formulation. Commonly used pharmaceuticallyacceptable excipients include water, magnesium stearate, starch,lactose, microcrystalline cellulose, stearic acid, sucrose, talc,silicon dioxide, gelatin, acacia and dibasic calcium phosphate(Baldrick, P. (2000) Regul. Toxicol. Pharmacol. October 32(2):210.)Excipients are combined with active ingredients for example to enhanceappearance, improve stability, aid processing or aid disintegrationafter administration, but many other excipient functions are known inthe art that can be applied to SAMe oral dosage forms. Classes ofexcipients which are often used and suitable for use in the presentinvention include but are not limited to, natural, modified-natural orsynthetic mono-, oligo- or polysaccharides where oligo- andpolysaccharides may or may not be physically or chemically crosslinked;natural, modified-natural or synthetic mono-, oligo- and polypeptides orproteins where oligo- and polypeptides and proteins may or may not bephysically or chemically crosslinked; synthetic oligomers and polymersthat may or may not be physically or chemically crosslinked; monomeric,hydrophobic, hydrophilic or amphoteric organic molecules; inorganicsalts or metals; and combinations thereof. Accordingly, SAMe may becombined with any excipient(s) known in the art that allows tailoringits performance during manufacturing as well as its in vitro and in vivoperformance. Many of these excipients may be utilized to tailor thedissolution profiles of SAMe formulations.

Disintegrants

Disintegrants are added to non-parenteral formulations to induce breakupof the product or dosage form (i.e. tablet or capsule) when it comes incontact with aqueous fluid in order to help release the drug. Theobjectives behind addition of disintegrants are to increase surface areaof the product fragments and to overcome cohesive forces that keep theseparticles together in a formulation. They do this by promoting wettingand swelling of the dosage form so that it breaks up in thegastrointestinal tract. Some binders such as starch and cellulose alsoact as disintegrants. Other disintegrants are clays, cellulosederivatives, algins, gums and crosslinked polymers. Another group ofdisintegrants called “super-disintegrants” are often utilized. Thesematerials are effective at low (2-5%) concentrations.“Super-disintegrants” which may be suitable for use in the presentinvention include, but are not limited to, sodium starch glycolate(SSG), croscarmellose sodium or crosprovidone.

The invention therefore also relates to compositions comprising SAMe andone or more disintegrants or “super-disintegrants” which improve thepharmacokinetic profile of SAMe in vivo.

Binders

The binding material which holds the bulk of the product together andalso helps maintain the product in a desired shape is known as a“binder” or “granulator”. Binders suitable for use in the presentinvention are exemplified by, but are not limited to, sugars, gelatin,gums, microcrystalline cellulose and modified celluloses, waxes orsynthetic polymers like polyethylene glycol or polyvinyl pyrrolidone.

Some embodiments of the invention may include improved pharmacokineticcompositions comprising SAMe and one or more binders.

Lubricants

Additional excipients often utilized in product formulations arelubricants. These are substances which aid in the manufacturing processas they help minimize clumping of the products and also help releasethem from the manufacturing machinery. The most common “lubricant” usedfor oral formulations is magnesium stearate; however, other commonlyused product lubricants include talc, calcium stearate, stearic acid(stearin), hydrogenated vegetable oils, sodium benzoate, leucine,carbowax 4000 and sodium stearyl fumarate all of which may be suitablefor use in the present invention.

Further exemplary embodiments of the invention also relate to improvedpharmacokinetic compositions comprising SAMe and one or more lubricants.

Glidants

Glidants also referred to as “flow-aids”, help to keep the powder makingup the products flowing as the products are being made, stopping themfrom forming lumps. Examples of commonly used glidants which may besuitable for use in the invention include colloidal silicon dioxide,talc, calcium silicate and magnesium silicate.

Additional embodiments of the invention relate to improvedpharmacokinetic compositions comprising SAMe and one or more glidants.

Coatings

An outer coating is typically applied to oral dosage forms in order tomask taste, odor or color; provide physical or chemical protection forthe active ingredient/drug; control the release of the active ingredientfrom the formulation; protect the active ingredient from the harshenvironment of the stomach (i.e. enteric coating); or protect thesubject from unwanted gastrointestinal side effects. Prior to applyingthe external coating, a seal coating may first be applied. Seal coatingsact to smooth the product surfaces, enhance the adherence of the final,outer coat and/or to protect the active ingredient from prematuredegradation. The type and/or thickness of the seal coat or the finalcoating(s) may be varied in order to alter product characteristics, suchas dissolution. Typically, the external or functional coatings aretargeted to be about 4-10% by weight and seal coats are targeted to beabout 1-5%, preferably about 2%, by weight. Seal coats are generallythought of as “non-functional” in that they are not utilized to controltiming or placement of release of the active ingredient; however, it isconsidered that certain seal coatings may act as such “functional”coatings. For the purpose of the present invention, “functionalcoatings” are intended to include enteric coatings, time-releasecoatings, pH-dependent coatings or other which control the timing orplacement of release of the active ingredient. In some exemplifiedembodiments of the invention, the one or more separate coatings orlayers of the functional coating together constitute 5% or less of thetotal dosage form targeted by weight. In preferred embodiments thefunctional coating is an enteric coating and even more preferably theenteric coating is about 3-4% targeted by weight.

Some embodiments of the invention may include improved pharmacokineticcompositions comprising SAMe and one or more coatings which alter thepharmacokinetic parameters of exogenous SAMe. Other embodiments of theinvention may include improved pharmacokinetic compositions comprisingSAMe and one or more coatings which alter the in vitro dissolutionprofile of SAMe. Specific exemplified embodiments of the inventioninclude improved pharmacokinetic compositions comprising SAMe and one ormore coatings which result in a dissolution profile exhibiting about25-80% SAMe released after one hour of incubation in the buffer phase;more preferably about 30-70% SAMe released within one hour of incubationin the buffer phase; and even more preferably, about 30-60% SAMereleased within one hour of incubation in the buffer phase.

The suitability of a particular excipient, such as, for example, a“matrix material”, “disintegrant”, “super-disintegrant” “binder”,“lubricant”, “glidant”, or “coating” may be identified by analyzing thein vivo pharmacokinetics of formulations containing the excipient andSAMe. Alternatively, in vitro analysis of one or more excipients using aseries of standard in vitro techniques which are well known in the artmay be used to pre-screen excipients and ultimately provide a means topredict in vivo pharmacokinetic profiles. Furthermore, the use ofreferences in the art may also provide insight into potentially suitablepharmaceutically or nutritionally acceptable excipients (such as a“matrix material”, “disintegrant”, “binder”, “lubricant”, “glidant”, or“coating”) for use in the present invention. Preferably, in vitroanalysis of one or more excipients using dissolution studies conductedwith a buffer pH of less than 6.8 may be used to pre-screen excipientsand ultimately provide a means to predict in vivo pharmacokineticprofiles.

Processing Methods and Parameters

Processing methods and/or parameters which may be modified in order toimprove the pharmacokinetic profile and/or alter the dissolution profileof SAMe formulations include but are not limited to: relative humidity,temperature, drying time and other environmental parameters.

Some exemplary embodiments of the invention are generated under lowhumidity conditions, typically less than or equal to about 35%, andpreferably less than or equal to about 15-25%, and more preferably lessthan or equal to about 10%. Other exemplary embodiments of the inventionare generated under manufacturing conditions wherein the temperature ismaintained between about 15-35° C. Other exemplary embodiments of theinvention are manufactured using a drying time of about 4-24 hours.Additional embodiments of the invention make use of SAMe compositionscomprising low water content. “Low water content” is preferably thoseformulations containing less than or equal to about 5% water, morepreferably less than or equal to about 3.5% water and even morepreferably less than or equal to about 1.5% water. In one way, watercontent is altered by controlling the relative humidity during themanufacturing process.

Exemplary embodiments of the invention relate to compositions comprisingexogenous SAMe and one or more suitable excipients and/or processingparameters which improve the pharmacokinetic profile of SAMe in vivo. Insome embodiments, the improved pharmacokinetic profile is identified byan average C_(max) of SAMe of at least about 1800 ng/mL, at least about1900 ng/mL or at least about 2000 ng/mL, when tested at a 1600 mg doseof SAMe ion; an AUC of at least about 7500 ng·h/mL, 8000 ng·h/mL, 8500ng·h/mL, or 9000 ng·h/mL when tested at a 1600 mg dose of SAMe ion; aT_(max) or C_(max) with reduced variation; a reduced effective dose, orcombinations thereof. In some preferred embodiments, the improvedpharmacokinetic profile is an average C_(max) of SAMe of at least 1800ng/mL for a 1600 mg dose of SAMe ion. In some embodiments, the improvedpharmacokinetic profile is identified by an average C_(max) of SAMe ofat least about 800 ng/mL, 825 ng/mL, 850 ng/mL, 875 ng/mL, 900 ng/mL, atleast about 950 ng/mL or at least about 1000 ng/mL, when tested at a 800mg dose of SAMe ion; an AUC of at least about 3000 ng·h/mL, 3250ng·h/mL, 3500 ng·h/mL, or 3750 ng·h/mL when tested at a 800 mg dose ofSAMe ion; a T_(max) or C_(max) with reduced variation; a reducedeffective dose, or combinations thereof. In some embodiments, theimproved pharmacokinetic profile is identified by an average C_(max) ofSAMe of at least about 400 ng/mL, 425 ng/mL, 450 ng/mL, at least about475 ng/mL or at least about 500 ng/mL, when tested at a 400 mg dose ofSAMe ion; an AUC of at least about 1500 ng·h/mL, 1550 ng·h/mL, 1600ng·h/mL, or 1650 ng·h/mL when tested at a 400 mg dose of SAMe ion; aT_(max) or C_(max) with reduced variation; a reduced effective dose, orcombinations thereof. In some embodiments, the improved pharmacokineticprofile is identified by an average C_(max) of SAMe of at least about400 ng/mL, at least about 450 ng/mL or at least about 500 ng/mL, whentested at a 400 mg dose of SAMe ion; an AUC of at least about 1500ng·h/mL, 1550 ng·h/mL, 1600 ng·h/mL, or 1650 ng·h/mL when tested at a400 mg dose of SAMe ion; a T_(max) or C_(max) with reduced variation; areduced effective dose, or combinations thereof. The improvedpharmacokinetic profile may also be identified by an average C_(max) ofSAMe of at least about 100 ng/mL per 100 mg dose of SAMe ion. Similarly,the improved pharmacokinetic profile may also be identified by an AUC ofabout 450 ng·h/mL per 100 mg dose of SAMe ion.

Dosing with Formulations Exhibiting Improved Pharmacokinetic Profiles ofSAMe

In some embodiments the improved pharmacokinetic SAMe formulations ofthe present invention are expected to be utilized to provide nutritionalsupport, or dietary supplement health improvements including, but notlimited to, mood improvement, joint health and liver function. In someexemplary embodiments the disorder is related to the dietary managementof a disease through additional supplementation of SAMe which cannot bereached through diet (i.e. a “medical food”.)

The improved pharmacokinetic SAMe formulations of the present inventionare suitable for therapeutic administration relating to a variety ofphysiological disorders and disease states, exemplified by, a mental orpsychiatric disorder (e.g. psychotic or non-psychotic mental disordersexemplified by depression and substance abuse disorders, respectively),a nervous system disease/disorder (e.g. a central nervous system diseaseexemplified by Alzheimer's), other neurological disease/disorders (e.g.headaches and sleep disorders), conditions associated with injury to thecentral nervous system, a liver disease/disorder (e.g. alcoholic liverdisease), a cancer (e.g. solid and blood-borne cancers), a jointdisease/disorder (e.g. arthritis), an inflammatory disease/disorder(e.g. ulcerative colitis), an autoimmune disease/disorder (e.g. systemiclupus erythematosis and rheumatoid arthritis), a degenerativedisease/disorder (e.g. Amyotrophic Lateral Sclerosis), a soft-tissuedisease/disorder (e.g. a fibromyalgia disorder), a paindisease/disorder, a genetic disorder related to hyper- orhypo-methylation, a gastrointestinal disease/disorder, a cardiovasculardisease/disorder, and a disorder induced in whole or in part byoxidative or free-radical damage.

Some embodiments of the present invention relate to therapeutic use ofthe exemplary compositions disclosed herein for treatment of a mental orpsychiatric disorder selected from the group consisting of anxietydisorders, depressive disorders, eating disorders, bipolar disorder,abuse disorders, dependence disorders, Axis II disorders, and psychosis.In some exemplary embodiments, the mental or psychiatric disorder is ananxiety disorder selected from the group consisting of generalizedanxiety disorder, posttraumatic stress disorder, social anxietydisorder, panic disorder, Schizophrenia and obsessive compulsivedisorder. In some exemplary embodiments, the mental or psychiatricdisorder is a depressive disorder selected from the group consisting ofmajor depressive disorder, multi-infarct dementia, minor depression,postpartum or late-life depression (and the like), Parkinson'sdepression, HIV-associated depression, brief recurrent depression,dysthymia or depression NOS (Not Otherwise Specified). In some exemplaryembodiments, the mental or psychiatric disorder is an eating disorderselected from the group consisting of bulimia nervosa, anorexia nervosa,binge eating disorder, obesity, or eating disorder NOS. In someexemplary embodiments, the mental or psychiatric disorder is bipolardisorder, an abuse disorder or a dependence disorder, including abuseof, or dependence on, alcohol, nicotine, cocaine, codeine, oxycodone,hydrocodone or other opiates. In some exemplary embodiments, the mentalor psychiatric disorder is an Axis II disorder selected from borderlinepersonality disorder.

In some exemplary embodiments, the disorder is a comorbid disorder, suchas comorbid depression arising in a subject who is undergoing treatmentfor one or more diseases or disorders such as but not limited to,cancer, Parkinson's and HIV. In certain embodiments the comorbiddisorder is caused by one or more therapies being utilized to treat saidone or more diseases or disorders.

In some exemplary embodiments, the disorder is a nervous systemdisorder, including a central nervous system (CNS) disorder such asParkinson's disease (and associated Parkinson's depression), Alzheimer'sdisease, Angelman Syndrome (genetic disorder), Multiple Sclerosis (MS)and pre-dementia and/or cognitive impairment.

In some exemplary embodiments, the disorder is a result of an injury tothe CNS such as spinal cord injury or brain damage, memory loss,cognitive impairment and/or learning disability.

In some exemplary embodiments, the disorder is a liver disorder selectedfrom the group consisting of alcoholic liver disease, fatty liverdisease (non-alcoholic) hepatitis (both viral and non-viral), livercancer, oxidative liver disease, HISS-dependent insulin resistance,cholestasis and cirrhosis.

In some exemplary embodiments, the disorder is a cancer selected fromthe group consisting of cancers occurring in one or more of the liver,colon, rectum, ovaries, urethra, testicles, bladder, breast, stomach,esophagus, pancreas, head and neck, lung, blood, skin (such as actinickeratosis, basal cell cancer, superficial basal cell cancer, squamouscell cancer, and melanoma) and adenocarcinomas.

In some exemplary embodiments, the disorder is a joint disorder such as,for example, arthritis and osteoarthritis.

In some exemplary embodiments, the disorder is an inflammatory disorderselected from the group comprising systemic lupus erythematosis, Reye'ssyndrome, rheumatic fever, allergic rhinitis, myasthenia gravis,temporal arteritis, vasculitis, psoriasis, atopic dermatitis, rosacea,eczema, alopecia universalis, scleroderma, pemphigus, contactdermatitis, ankylosing spondylitis, dermatomyositis, polymyositis,celiac sprue, Guillain-Barré syndrome, multi-infarct dementia,post-cerebral vascular accident reperfusion damage, Addison's disease,Hashimoto's thyroiditis, asthma, upper respiratory inflammationsymptoms, chronic bronchitis, atherosclerosis, pernicious anemia,autoimmune hepatitis, prostatitis, pelvic inflammatory disease,Goodpasture's syndrome, Wegener's granulomatosis, chronic nephritis,Sjogrens syndrome, or allergic conjunctivitis.

In some exemplary embodiments, the disorder is a gastrointestinaldisorder such as inflammatory bowel disease (IBD), Crohn's disease orulcerative colitis (UC).

In some exemplary embodiments, the disorder is a soft tissue diseasesuch as fibromyalgia.

In some exemplary embodiments, the disorder is a pain disorder such asfibromyalgia, chronic headaches, shingles, reflex sympathetic dystrophyand polyneuropathy.

In some exemplary embodiments, the disorder is a cardiovascular disorderwhich is related to hyper- or hypo-homocysteinemia such as coronaryheart disease, stroke, peripheral vascular disease and atheroscleroticdisease.

In some exemplary embodiments, the disorder is related to a genetic ormedical condition related to a deficiency of the methylation pathwaysuch as methylenetetrahydrofolate reductase deficiency.

In some exemplary embodiments, the etiology of the disorder may includeoxidative or free-radical damage, and is selected from the groupcomprising chronic fatigue syndrome, temporal arteritis, vasculitis,multi-infarct dementia, chronic emphysema, ischemia-reperfusion injury,chronic nephritis or vascular depression.

In some embodiments the improved pharmacokinetic SAMe formulations ofthe present invention are expected to exhibit an improved side effectprofile when utilized to provide nutritional support or dietarysupplement health improvements or for therapeutic administrationrelating to a variety of physiological disorders and diseases such asthose listed above. Side effect which may be improved upon include, butare not limited to, gastrointestinal (e.g. nausea, loose stools orconstipation); insomnia; restlessness; sexual dysfunction; lowerhomocysteine levels; cardiovascular side effects (e.g. heartpalpitations); nervousness; loss of appetite; dry mouth; dizziness andheadaches.

The improved PK SAMe formulations of the invention provide an increasein SAMe plasma levels which are significantly improved in comparison toconventional non-parenteral SAMe dosage forms. There are many potentialbenefits of this increase in exposure of SAMe to the body includingreaching a more therapeutically effective amount of SAMe (such as thoseresulting from parenteral SAMe administration) which may be necessary toachieve and/or improve a clinical benefit to one or more of thediseases/disorders listed above. Thus, in some embodiments of theinvention provided are SAMe formulations which exhibit rapid-onset oftreatment for one or more of the diseases/disorders listed above.

In some embodiments the improved pharmacokinetic SAMe formulations ofthe present invention exhibit average SAMe AUC values of at least about7500 ng·h/mL per day and/or an average C_(max) of about 1800 ng/mL perday. In other embodiments, average SAMe AUC values of at least about7500 ng·h/mL per day and/or an average C_(max) of about 1800 ng/mL perday lead to improved efficacy in comparison to treatment withconventional exogenous SAMe formulations. These improved C_(max) and/orAUC values may be achieved using a number of dosing regimens and thusmay result from a SAMe ion dose which is less than 1600 mg and may be aresult of administration of one or multiple SAMe oral dosage forms overthe course of 24 hours.

Suitable subjects for dosing according to the methods and compositionsof the invention include warm-blooded mammals such as humans, domesticor exotic animals or livestock; domesticated avian subjects such aschickens and ducks; and laboratory animals suitable for research use.When used for treating a disease or disorder in a subject, varioussymptoms of specific physiological disorders and disease states arecontemplated as being treatable within the context of the presentinvention and details of which are set forth below. However, it is to berecognized that the understanding of various disease states by those ofskill in the art is not static and this is the same for performancevariables related to nutritional supplementation. Thus, though thedescription above is intended to be illustrative of the variousdisorders, disease states, symptoms or performance variables that may betreated using the improved pharmacokinetic SAMe formulations accordingto the present invention, a person skilled in these arts will beexpected to apply such knowledge.

Dosing with Multiple Dosing Units

Some exemplary embodiments of the present invention relate to treatmentof and/or prophylaxis of one or more diseases in a subject wherein thetreatment of and/or prophylaxis of one or more diseases and/or disorderscomprises administering to the subject a physiologically effectivedosage comprising S-adenosyl methionine (SAMe), or a proprietary saltthereof, which exhibits an improved pharmacokinetic profile in vivo.

In some exemplary embodiments, the dosing schedule may be dividedbetween multiple daily doses. Multiple daily doses need not be identicaland may comprise one or more dosage forms in combination. In someexemplary embodiments, the improved pharmacokinetic SAMe may be dividedinto two or more daily doses. Each dose may be administered as a singledosage unit exemplified by a single tablet, capsule or caplet, oralternatively may be divided into multiple dosage units. In someembodiments, a twice-daily dose of from about 100 to about 1600 mg ofSAMe ion per dose may be divided into one to four dosage units of fromabout 100 to about 800 mg of SAMe ion per unit. In each case, the formof the dosage unit may be a capsule, a tablet, a caplet or an extendedrelease dosage unit and the like. As mentioned previously, SAMe API issupplied as a molecular entity comprising an ion along with severalcounter-ions and when referring to SAMe dosing, it is currently acceptedin the art that the numerical dose (usually in milligrams) refers to theamount of SAMe ion which is administered. Therefore, a “400 mg SAMetablet” is referring to a tablet which contains 400 mg of the SAMe ion.

Conventional SAMe dosing generally administers up to 1600 mg of SAMe ionper day (800 mg twice daily). Tablets are most often availablecommercially in 200 mg and 400 mg (SAMe ion) doses which requiresubjects to ingest 4-8 tablets per day. This is inconvenient withrespect to the amount of time needed as well as the potential error inconsistent dosing (i.e. if a dose is missed). The present invention hasidentified novel compositions and methods which reduce the effectivedose of SAMe and/or eliminate the need to dose twice or more daily. Byimproving the pharmacokinetic profile, a new method of SAMe therapy isavailable which potentially lowers the amount of SAMe dose required toelicit an effective response by providing compositions comprising one ormore suitable excipients or final dosage form characteristics whicheither increase the average C_(max) or AUC or reduce the variation inpharmacokinetic parameters. These exemplary “low dose” formulations mayprovide a lower daily pill count which is beneficial to those takingSAMe as it will reduce the time, cost and inconvenience ofself-administering large doses. It is expected that lower dosing willalso lead to an improved side effect profile when compared to doses ofother SAMe products which exhibit similar PK profiles. Thus in someembodiments, provided are SAMe compositions which improve the sideeffect profile of exogenous SAMe formulations.

In additional exemplary embodiments, the effective dose is administeredon a once a day basis. In some embodiments, the once a day dose may beadministered in a single dosage unit exemplified by, a single tablet,capsule, or caplet. In other exemplary embodiments, the single dose maybe administered as multiple tablets, capsules or caplets taken at onetime. In some embodiments, for instance, a dosage of about 400 to 3200mg of SAMe per day may be divided into two, three, four or more tablets,capsules or caplets of about 100 to 1600 mg of SAMe per unit dose. Insome preferred embodiments, the daily dose may comprise two, three orfour tablets, capsules or caplets of about 100 to 800 mg of SAMe perdose. Suitable dosage regimens included are: four units of about 200 or400 mg SAMe per unit; three units of about 100, 150, 200, 300, 400, 600,800 or 1,000 mg of SAMe per unit; two units of about 200, 400, 800 or1600 mg per unit.

In certain embodiments of the invention, the effect of once a day dosingis believed to result in the most consistent pharmacokinetic parametermeasurements, specifically the C_(max) and T_(max). The “more reliable”pharmacokinetic profile which results from once a day dosing of theseformulations allows for improved knowledge of dosing by the medicalpractitioner as well as improved pharmacokinetic profiles with regard tothe time of day when the subjects experience the highest systemicexposure of SAMe (C_(max)) which is anticipated to give rise to animprovement in the side effect profiles of these subjects.Traditionally, SAMe dosing is administered twice daily on a fastedstomach for the initial dose. Because subjects are fasting, the firstdose is generally given early in the morning (i.e. 8:00 am) prior tofood intake. The second dose is then given 8 hours later (i.e. 4:00 pm),typically to a non-fasted stomach. A strong “food affect” is routinelyobserved which is characterized by a delay in the T_(max) such that theC_(max) is not achieved until normal sleep hours (often 11:00 pm to 2:00am). Insomnia and other sleep-related events are often reported for manyconventional SAMe formulations and this may occur because of thenighttime C_(max) caused by the second dose and the correspondingstimulatory effects of SAMe. In contrast, once a day dosing in themorning using formulations of the invention results in the delivery ofequivalent or higher total daily amounts of SAMe (i.e. similar AUCvalues) yet with the potential for reduced insomnia side effects likelybecause the T_(max) and C_(max) are achieved during normal waking hours.

Therefore some exemplary embodiments of the present invention relate tocompositions comprising SAMe in combination with at least one suitableexcipient or product characteristic, wherein said compositions areadministered using a once a day dosing regimen. In some embodiments, theSAMe compositions are administered pre-prandially, e.g. beforebreakfast, thus on a fasted stomach. Certain embodiments also relate tocompositions comprising SAMe in combination with at least one suitableexcipient or product characteristic, wherein said compositions exhibit areduced effective dose of the administered physiologically effectivedosage of SAMe. Preferably, such compositions achieve an enhancedpharmacokinetic profile such as, for example, an average maximum SAMeblood plasma concentration (average C_(max)) of at least about 100 ng/mLper 100 mg of SAMe ion when administered in vivo.

Combinations of Same with Other Active Ingredients

Some exemplary embodiments of the present invention relate tocombinations of SAMe with one or more active ingredients that arecommonly prescribed or used for treating and/or prophylaxis in a subjecta disease or disorder selected from the group consisting of, but notlimited to, a mental or psychiatric disorder (e.g. psychotic ornon-psychotic mental disorders such as depression and substance abusedisorders, respectively), a nervous system disease/disorder (e.g. acentral nervous system disease such as Alzheimer's), other neurologicaldisease/disorders (e.g. headaches and sleep disorders), conditionsassociated with injury to the central nervous system, a liverdisease/disorder (e.g. alcoholic liver disease), a cancer (e.g. solidand blood-borne cancers), a joint disease/disorder (e.g. arthritis), aninflammatory disease/disorder (e.g. ulcerative colitis), an autoimmunedisease/disorder (e.g. systemic lupus erythematosis and rheumatoidarthritis), a degenerative disease/disorder (e.g. Amyotrophic LateralSclerosis), a soft-tissue disease/disorder (e.g. a fibromyalgiadisorder), a pain disease/disorder, a genetic disorder related to hyperor hypo methylation, a gastrointestinal disease/disorder, acardiovascular disease/disorder, and a disorder induced in whole or inpart by oxidative or free-radical damage, comprising administering tosaid subject an exemplary composition of the present invention whichimproves the pharmacokinetic profile of a physiologically effectiveamount of exogenous SAMe.

Additionally, combinations of SAMe with one or more active ingredientsas detailed above may act to ameliorate the side effects associated withsaid one or more active ingredients. Combinations with SAMe may beco-administered or taken separately and need not be taken at the sametime.

Other embodiments of the present invention relate to combinations ofSAMe with one or more active ingredients that are commonly prescribed orused for treatment of and/or prophylaxis of mental or psychiatricdisorders in a subject include, but are not limited to, tricyclicantidepressants (TCAs), tetracyclic antidepressants, aminoketones,phenylpiperazines, selective serotonin reuptake inhibitors (SSRIs),monoamine oxidase inhibitors (MAOIs), serotonin-norepinephrine reuptakeinhibitors (SNRIs), norepinephrine-serotonin reuptake inhibitors(NSRIs), dopamine reuptake inhibitors, norepinephrine-dopamine reuptakeinhibitors, norepinephrine reuptake inhibitors, selective serotoninreuptake enhancers, noradrenergic and serotonin specificantidepressants, substance P receptor antagonists, neurokinin receptorantagonists such as saredutant, corticotrophin release factorantagonists such as mifepristone, atypical antipsychotics such asaripiprazole, commonly used antidepressant augmenters such as lithium,triple reuptake inhibitors and the like.

Some embodiments of the present invention relate to combinations of SAMewith one or more device therapies that are commonly prescribed or usedfor treatment of and/or prophylaxis of mental or psychiatric disordersin a subject include, but not limited to ECT (electro convulsivetherapy) and electric shock therapy.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a nervous system disease/disorderin a subject include, but are not limited to anticonvulsants such aspregabalin, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)receptor antagonists, methylphosphonate (NMPA) receptor antagonists,histamine receptor antagonists, nitric oxide (NO) modulators, glutamatereceptor antagonists, acetylcholinesterase inhibitors, dopamineagonists, N-methyl-d-aspartate (NMDA) receptor antagonists such asmemantine, cholinesterase inhibitors such as donepezil,neuroprotectants, nootropic agents, CNS modulators, antiamyloidogenics.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a liver disorder in a subjectinclude, but are not limited to, antiviral medication such as alphainterferon, ribavirin, lamivudine, steroids, antibiotics and zincacetate.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a cancer in a subject include,but are not limited to, chemotherapeutic agents, drug resistancemodulators, monoclonal antibodies, cytokines (e.g. interferons andinterleukins), immunocytokines, growth factors, chemoprotectants,vaccines and other biological response modifiers.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a joint or inflammatorydisease/disorder in a subject include, but are not limited to,analgesics, non-steroidal anti-inflammatory drug compounds (NSAID),disease-modifying antirheumatic drugs (DMARDs), corticosteroids,anakinra (an interleukin-1 receptor antagonist), COX-2 inhibition,gamma-aminobutyric acid-B (GABAB) receptor agonists, such as baclofen,GABAA potentiating drugs, such as the benzodiazepines tumor necrosisfactor (TNF)-inhibiting drugs, and other drugs that modify the immuneresponse (immunosuppressive drugs).

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of an autoimmune disease/disorder ina subject include, but are not limited to, DMARDs, corticosteroids,anakinra (an interleukin-1 receptor antagonist), TNF-inhibiting drugs,and other drugs that modify the immune response (immunosuppressivedrugs).

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a degenerative disease/disorderin a subject include, but are not limited to, NSAIDs, COX-2 inhibition,GABAB receptor agonists, such as baclofen, and GABAA potentiating drugs,such as the benzodiazepines.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a soft tissue disease/disorder ina subject include, but are not limited to, milnacipram, pregabalin,SNRIs, NSRIs, muscle relaxers, sedatives, painkillers, and NSAIDs.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a genetic disease/disorderrelated to hyper or hypo methylation in a subject include, but are notlimited to methionine, MTA (5′-deoxy-5′-(methylthio)adenosine), andother SAMe metabolites.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a gastrointestinaldisease/disorder in a subject include, but are not limited to,5-Aminosalicylic acid (5-ASA) medications, Corticosteroids (prednisone),immunomodulatory medications such as Azathioprine (Immuran),6-Mercaptopurine (6-MP), Methotrexate and Cyclosporine (Sandimmune),commonly used antibiotics such as Metronidazole (Flagyl) andCiprofloxacin (Cipro) and biologic agents such as Infliximab (Remicade).

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a cardiovascular disease/disorderin a subject include, but are not limited to, statins,angiotensin-converting enzyme (ACE) inhibitors, ASA, SAMe break downproducts such as methionine, MTA and folate, cardioprotectants,vasoprotectants, coagulation inhibitors.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a disorder induced in whole or inpart by oxidative or free-radical damage including, but are not limitedto, antioxidants such as Vitamin A, Vitamin C, Vitamin E, polyphenols,flavonoids, selenium, carotenoids.

Some embodiments of the present invention relate to combinations of SAMewith one or more active ingredients that are commonly prescribed or usedfor treatment of and/or prophylaxis of a disorder induced in whole or inpart by damage to the central nervous system such as brain injury orspinal cord injury including, but not limited to, neuroprotectants,nootropic agents, CNS modulators, analgesics, muscle relaxants,apoptosis inhibitors, bone modulators, antioxidants.

Some embodiments of the present invention relate to combinations of SAMewith methionine, MTA, folate, vitamin B6 and/or B12 as they are eachcorrelated with lowering homocysteine production. Therefore, it isconsidered that combining SAMe with methionine, MTA, folate, methylfolate, vitamin B6 and/or B12 may result in increased supplementation ofSAMe by enhancing the body's natural ability to make SAMe while at thesame time supplementing SAMe with exogenous SAMe exhibiting an improvedpharmacokinetic profile.

In some embodiments, an exemplary improved pharmacokinetic SAMe dosageform according to the invention may be included in a kit with a separatedosage form containing at least one other active ingredient, exemplifiedby one or more compounds suitable for the treatment of or commonlyprescribed or used for the treating and/or prophylaxis in a subject adisease or disorder selected from the group consisting of, but notlimited to, a mental or psychiatric disorder (e.g. psychotic ornon-psychotic mental disorders such as depression and substance abusedisorders, respectively), a nervous system disease/disorder (e.g. acentral nervous system disease such as Alzheimer's), other neurologicaldisease/disorders (e.g. headaches and sleep disorders), conditionsassociated with injury to the central nervous system, a liverdisease/disorder (e.g. alcoholic liver disease), a cancer (e.g. solidand blood-borne cancers), a joint disease/disorder (e.g. arthritis), aninflammatory disease/disorder (e.g. ulcerative colitis), an autoimmunedisease/disorder (e.g. systemic lupus erythematosis and rheumatoidarthritis), a degenerative disease/disorder (e.g. Amyotrophic LateralSclerosis), a soft-tissue disease/disorder (e.g. a fibromyalgiadisorder), a pain disease/disorder, a genetic disorder related to hyperor hypo methylation, a gastrointestinal disease/disorder, acardiovascular disease/disorder, and a disorder induced in whole or inpart by oxidative or free-radical damage, comprising administering tosaid subject an exemplary composition of the present invention whichimproves the pharmacokinetic profile of a physiologically effectiveamount of exogenous SAMe.

In some embodiments, an exemplary improved pharmacokinetic SAMe dosageform according to the invention may be included in a kit with a separatediagnostic agent or tool exemplified by one or more agents/toolssuitable for use as part of a diagnostic test. In certain embodimentsthe diagnostic agent or tool is used as part of a test for measuring thelevels of one or more biomarkers.

In addition to combinations of SAMe with the one or more additionalingredients exemplified above or methionine, MTA, folate, vitamin B6and/or B12, administration of the exemplary improved pharmacokineticSAMe formulations of the invention may also augment the effects of otherdrugs or nutritional supplements being taken by the subject. Thus, someexemplary embodiments of the present invention relate to combinations ofSAMe with drugs or nutritional compounds already employed for treatingother diseases for increasing the activity of said drugs or nutritionalcompounds.

The present invention is further described by the following examples.These examples, while illustrating certain specific aspects of theinvention, should not be considered to limit or circumscribe the scopeof the disclosed invention.

EXAMPLES Example 1 SAMe Formulations of the Invention Generate C_(max)Values Significantly Greater than those from Commercially Available SAMe

In order to identify and optimize processing components, methods andparameters that impart product characteristics which result in improvedpharmacokinetic profiles, SAMe formulations comprising variousexcipients and prepared with certain final dosage form characteristicswere compared with a commercially available SAMe formulation. In thisexample, a commercially available S-adenosyl methionine tosylatedisulfate formulation was utilized as the control SAMe dosage form.

SAMe formulations of the invention were generated using standardprocedures for making tablets and comprise the following:

Ingredient (MSI-43) % (w/w) SAMe Disulfate Tosylate 76.8Microcrystalline Cellulose 113 7.6 Microcrystalline Cellulose 112 9.1Disintegrant 5 Colloidal Silicon Dioxide 0.5 Magnesium Stearate 1.0

In order to improve the compressibility of SAMe, a dry granulationprocedure (“slugging”) was employed. The above ingredients were mixedand pressed through a mesh screen. Slugs were made on a Manesty SP pressfitted with 16/32 tooling to a hardness of 2-4 kPa. Slugs were thenmilled on an Erweka Dry Granulator and pressed through a mesh screen.The final mixture was then compressed to a hardness of 12-17 kPa on aStokes DS3 semi automatic press using an elongated oval die. Humiditywas maintained at below 30% and temperature was maintained at 20-25° C.during the entire manufacturing process. The granules demonstrated goodflow properties and no sticking or picking during compression.

Prior to applying an enteric coating to these tablets, a seal coat wasfirst applied in order to improve the tablet surface properties. Astirred 12% suspension of a commercially available seal coat in purifiedwater was applied to the uncoated SAMe tablets in an Aeromatic CoatingColumn using 55° C. inlet air temperature and 4-6 g/min spray rate until2% weight gain was achieved.

A plasticized 80% solid (w/w) commercially available enteric coatdesigned to dissolve at pH 5.5 was then applied to the tabletformulations such that they would remain intact within the stomach. Theenteric coating was generated using a stirred 30% aqueous suspension(56% of final coating suspension by weight), plasticizer 20% aqueoussuspension (7% of final coating suspension by weight), triethyl citrate(2% of final coating suspension by weight) and purified water (35% offinal coating suspension by weight). These components were mixed andthen applied to the seal coated SAMe tablets in an Aeromatic CoatingColumn using 55° C. inlet air temperature and 4-6 g/min spray rate.

The manufacturing process produced dosage form characteristics whichwhen combined with the specific combinations of SAMe and excipients gaverise to the exemplary SAMe formulation of the invention. Tablets wereexamined based on several in vitro test criteria including thefollowing:

-   -   Hardness: average range of 16 with variance (SDEV squared) of 3    -   Average weight of tablets: 1100 mg (target 400 mg SAMe ion with        standard pharmaceutical variance)    -   Average size: 9.2 mm×19.1 mm    -   Exterior Coating thickness: 6.4% weight gain    -   Water Content: 2.2% w/w

Once the final tablets of the invention were generated andcharacterized, they were administered to healthy volunteers in order tocompare their in vivo pharmacokinetic profiles to those of acommercially available and routinely used SAMe tosylate disulfateformulation.

For the control group, seven healthy, male volunteers who had fastedwere given a single dose of 1600 mg of the commercially availableS-adenosyl methionine tosylate disulfate formulation. Similarly, for thetest group, nine healthy and fasted male volunteers were administered1600 mg of the SAMe formulation of the invention (“MSI-43”) given as asingle dose. The resulting pharmacokinetic profiles were studied bymeasuring the presence of SAMe in plasma at various time points afteradministration.

The graph in FIG. 1 shows the average SAMe plasma concentration curvesfor the aforementioned subjects each of whom received either a single1600 mg dose of the commercially available S-adenosyl methioninetosylate disulfate formulation or a single 1600 mg dose of the novelSAMe formulation of the invention, MSI-43. For the commerciallyavailable SAMe formulation, an average C_(max) of about 900 ng/mL wasreached within approximately 3-4 hours of administration. As can be seenin the graph, and what was particularly of note here was that subjectswho received a single dose of the SAMe formulation of the inventionrecorded an average C_(max) of approximately 4000 ng/mL within less thanabout 3 hours of administration. These results clearly show that SAMeformulations of the present invention provide a significant improvementto the pharmacokinetic profile of SAMe once administered and lead toincreases in SAMe C_(max) and AUC values never disclosed before in theprior art outside of injectable forms.

Example 2 Targeted Dissolution “Window” Results in SuperiorPharmacokinetic Profiles In Vivo

In this example, additional SAMe formulations were made, which haddiffering amounts of disintegrants, coating thickness, and weregenerated under various processing parameters in order to compare theirin vivo performance and in vitro dissolution profiles. Each formulationcomprised:

Ingredient % (w/w) SAMe Disulfate Tosylate 76.8 MicrocrystallineCellulose 113 7.5-9   Microcrystalline Cellulose 112 11-13.5Disintegrant 0-10  Colloidal Silicon Dioxide 0.5 Magnesium Stearate 1.0The disintegrants used were sodium starch glycolate, croscarmellose orcrosprovidone. The percent of microcrystalline cellulose may varyaccording to the amount of disintegrant, such that the final % (w/w) ofthe formulation totals 100%. The tablets were manufactured according tothe processes as described in Example 1. The in vitro attributesdescribed in Examples 1 were also measured. In vivo human tests wereconducted as per described in Example 1. The following table provides asummary of the tablet characteristics and corresponding pharmacokineticattributes and focuses on those formulations which exhibit improvedpharmacokinetic profiles:

TABLE 1 IN VIVO PROFILE OF VARIOUS SAMe FORMULATIONS DOSED AT 1600 mgEnteric Average Coating Dissolution Cmax* RH/ Thickness Average at 30min (ng/mL AUC_(0-t){circumflex over ( )}{circumflex over( )}{circumflex over ( )} T/C at % at by hardness/ post of Tmax** (h *ng/mL tableting tableting weight % kP 6.8 pH SAMe) (hrs) of SAMe) MSI-0317 32 8.0 n/a 5 401 3 4361 MSI-09 24 34 8.0 12 4 1158 4 6929 Commercialn/a n/a n/a 34.1 10 765 5 5261 Commercial n/a n/a n/a 34.1 10 941 5 4504MSI-72 25 23 2.9 15.2 96 2067 3 10365 MSI-90 32 28 3.6 15.4 86 2561 414013 MSI- 33 2 3.7 15.0 78 1846 3 10440 104 MSI- 29 15 3.6 17.4 94 32235 13402 111 *Average Cmax was calculated by averaging all subjects ateach time point and then taking the maximum of the averages **Tmax isthe time point where Cmax* was determined {circumflex over( )}{circumflex over ( )}{circumflex over ( )}AUC is area under thecurve

Example 2a Targeted Dissolution With Different Excipients

In this example, additional SAMe formulations are made, which havediffering amounts of disintegrants and coating thickness, and differentlubricants. The formulations are generated under various processingparameters in order to compare their in vivo performance and in vitrodissolution profiles. Each formulation comprised:

Ingredient % (w/w) SAMe Disulfate Tosylate    60-80% MicrocrystallineCellulose 113  7-10 Microcrystalline Cellulose 112 10-15 Disintegrant 0-10 Colloidal Silicon Dioxide 0-1 Lubricant 0.1-2  The disintegrants used may be sodium starch glycolate, croscarmellose,crospovidone or some other suitable disintegrant. The percent ofmicrocrystalline cellulose may vary according to the amount ofdisintegrant, such that the final % (w/w) of the formulation totals100%.

The tablets are manufactured according to the processes as described inExamples 1 and 2. The in vitro attributes described in Examples 1 arealso measured. In vivo human tests are conducted as per described inExamples 1 and 2. The following table provides a summary of thelubricants that may be employed in the tablets according to theinvention:

Exemplary Lubricants Calcium stearate Stearic acid Sodium benzoateLeucine Sodium stearyl fumarate Talc

Example 3 In Vitro Dissolution Profiles as a Predictor of In VivoPerformance

Dissolution profiles are routinely used as an in vitro marker to confirmfinal product and/or tablet characteristics. This profiling is typicallyperformed according to USP standards and under conditions which mimicphysiological pH (pH 6.8).

The dissolution test method used is typically as follows:

-   -   USP Apparatus II operated at 100 RPM    -   Fluid Phase: 1 L USP Simulated Gastric Fluid without enzyme, pH        1.2, 37° C.    -   Aqueous Buffer Phase—1 L USP simulated Intestinal Fluid without        enzyme, pH 6.8, 37° C.    -   Tablets are exposed to the acid phase for two hours then        transferred to the Buffer Phase    -   Aliquots are drawn following exposure to the acid phase for 2        hours, then at prescribed intervals while in the buffer phase    -   Samples are diluted 1→10 with n/10 HCL    -   Drug concentration is determined spectrophotometrically at 258        mm

Eight different SAMe formulations were compared against the samecommercially available SAMe product as mentioned previously and thedissolution profiles obtained are represented in FIG. 2A. As shown inthe graph, the eight SAMe formulations of the invention all show rapiddissolution within 30 minutes of incubation in the aqueous buffer phasein comparison to the Commercially available SAMe (note this is between 2and 2.5 hours on the X-axis as all samples are exposed to the acid phasefor 2 hours prior to transfer and analysis in the buffer phase).Furthermore, the dissolution profiles of the eight SAMe formulations ofthe invention overlap such that it is difficult to differentiate betweenthese formulations and all appear to be equally “rapid”.

The SAMe formulations of the invention are coated with an entericcoating designed to dissolve in the region exiting the stomach andtherefore the present investigators designed a new profiling methodwhich would better mimic the conditions of the small intestine. Usingthis method, the buffer phase was adjusted to pH 6.0 rather than 6.8.

Surprisingly, the in vitro dissolution profiles generated under theseconditions identified a clear distinction between the eight formulationsof the invention as those that are either faster or slower to releasedrug in the earliest time points. The graph in FIG. 2B shows theseprofiles and clearly depicts MSI-111, MSI-90 and MSI-104 as‘fast-release’ formulations. As seen in the table in Example 2 above, invivo profiling of all of these formulations shows that thesefast-release formulations have very high C_(max) and AUC values andrepresent those formulations of the invention which exhibit improvedpharmacokinetic profiles of SAMe.

Also, when a commercially available rapid-dissolve SAMe product,“Commercial product 1”, was compared against various formulations of theinvention it was discovered that this extremely fast-dissolvingformulation did not exhibit the very high in vivo SAMe C_(max) and/orAUC values as exemplified and claimed in the present invention.Similarly, dissolution profiling of another commercially available SAMeproduct (“Commercial product 2”), which was utilized in the Examplesabove as well as the following Examples, showed that at pH 6.0 thedissolution of this product was “slow” in comparison to the improved PKSAMe formulations of the invention (a summary of these dissolutionprofiles is seen in FIG. 3). Therefore, surprisingly, there exists a“window” of dissolution that is neither too fast nor too slow and leadsto unexpectedly high levels of SAMe in the plasma. Provided herein isthus a novel in vitro method for identifying SAMe formulations whichexhibit improved pharmacokinetic profiles in vivo.

Example 4 Effect of Coating Thickness, Temperature and Relative Humidityon SAMe Pharmacokinetic Parameters

The in vivo profiles of multiple SAMe formulations of the inventioncomprising different excipients and/or dosage form characteristics werecompared. As seen in Table 2 below, the ‘thin-coated’ formulationsgenerated high C_(max) and AUC values. “Thin” coatings are meant toinclude those that are less than about 6% but more preferably targetedto about 4%. As seen in the table, relative humidity was maintainedbelow 30%. It was necessary to maintain this low humidity in order togenerate workable “thin” coated formulations. Temperature was maintainedat less than 35° C.

TABLE 2 IN VIVO PROFILE (ranked by individual's averages) dose 1600 mgSAMe ion Enteric Coating Thickness- Dissolution Cmax*AUC_(0-t){circumflex over ( )}{circumflex over ( )}{circumflex over ( )}T/C RH/ targeted average at 0.5 hrs (ng/mL (h * ng/ at % at average byhardness/ post of Tmax** mL of tableting tableting weight/g weight % kP6.8 pH SAMe) (hrs) SAMe) Humidity between 15-35% and “thin” coatings:MSI- 29 15 1.1 3.6 14.0 94 (84 at 3223 6 13402 111 1.5 hrs post pH 6.0)MSI-90 32 28 1.1 3.6 15.0 86 (78 at 2561 4 14013 1.5 hrs post pH 6.0)MSI-72 25 23 1.1 2.9 15.2 96 2067 3 10365 Humidity at or below 10%:MSI-43 22 4 1.1 6.4 16.0 70 4017 3 13168 MSI-77 22 0.6 1.1 6.5 17.4 832635 5 13927 MSI-78 22 4 1.1 6.7 16.0 78 2299 4 13642 MSI-79 22 4 1.16.9 15.4 54 2113 4 10250 MSI- 33 2 1.1 3.7 15.0 95 (67 at 1846 3 10440104 1.5 hrs post pH 6.0) MSI- 33 2 1.1 5.9 15.0 78 2190 3 9994 105*Average Cmax was calculated by averaging all subjects at each timepoint and then taking the maximum of the averages **Tmax is the timepoint where Cmax* was determined {circumflex over ( )}{circumflex over( )}{circumflex over ( )}AUC is area under the curve

When the functional (or enteric) coating thickness is reduced to around4%, improved PK SAMe formulations of the invention are generated underconditions where the humidity levels can be as high as 35%. However,surprisingly, the present investigators also discovered that improved PKSAMe formulations of the invention generated under very low humidityconditions (less than or equal to 10%) are less sensitive to thethickness of the outer coating and improved PK SAMe formulations of theinvention can be generated with functional coatings as thick as 9-10%.Therefore, the invention also relates to a method for making SAMeformulations with improved PK parameters by manufacturing saidformulations under conditions wherein the relative humidity is about 10%or less.

The graph in FIG. 4 shows the effect of relative humidity andtemperature on the glass transition temperature (Tg) of SAMe. As seen inthe graph, there is a preferred operating range of these processparameters when preparing SAMe formulations of the invention in order toyield a Tg greater than room temperature. The lowest humidity (under10%) is more preferable for those formulations where the coatingthickness remains above about 4%.

Example 5 SAMe Formulations of the Invention Result in a ReducedEffective Dose

SAMe formulations of the invention comprising different excipientsand/or dosage form characteristics were compared with a commerciallyavailable SAMe formulation at both comparative doses as well as at halfthe dose. In this example, the same commercially available S-adenosylmethionine tosylate disulfate formulation stated before was utilized asthe control SAMe dosage form.

A first SAMe formulation of the invention (termed, “MSI-72”) comprisingsodium starch glycolate (SSG), colloidal silicon dioxide and magnesiumstearate was generated using similar excipients and procedures asdescribed in Example 1. The specific MSI-72 formulation was as follows:

Ingredient % (w/w) SAMe Disulfate Tosylate 76.8 MicrocrystallineCellulose 113 7.6 Microcrystalline Cellulose 112 9.1 Sodium StarchGlycolate 5 Colloidal Silicon Dioxide 0.5 Magnesium Stearate 1.0

As detailed in Example 1, the same seal coat followed by an entericcoating designed to dissolve at pH 5.5 was applied to the tablets.

For comparison of the above MSI-72 SAMe formulation of the inventionwith a commercially available SAMe formulation, a single 1600 mg dose ofeither the commercially available S-adenosyl methionine tosylatedisulfate formulation or the MSI-72 SAMe formulation was given to twogroups of healthy and fasted, male volunteers (seven in each group). Asingle dose of 800 mg of the proprietary SAMe formulation was alsocompared in a third study group to the same 1600 mg dose of thecommercially available SAMe. The resulting pharmacokinetic profiles werestudied by measuring the presence of SAMe in plasma at various timepoints after administration.

The graph in FIG. 5A shows the average plasma SAMe concentration curvefor seven subjects who received a single 1600 mg dose of thecommercially available S-adenosyl methionine tosylate disulfateformulation in comparison to the average plasma SAMe concentration curvefor seven subjects who received 1600 mg of the SAMe formulation of thepresent invention (MSI-72) given as a single dose. As can be seen in thegraph, the average C_(max) for the proprietary SAMe formulation issignificantly increased in comparison to conventional SAMe therapy.

Moreover, the data presented in FIG. 5B represent the same 1600 mg doseof conventional, commercially available SAMe therapy as detailed above;however the MSI-72 SAMe formulation was dosed at only half of the dose.800 mg of MSI-72 was administered as a single dose to seven healthy andfasted, male volunteers and the presence of SAMe in plasma was measuredafter administration at the indicated time points. As can be seen in thegraph, the average C_(max) of the inventive formulation when using onlyhalf the dose is comparable to the full, 1600 mg dose of thecommercially available SAMe formulation.

Another SAMe formulation of the invention (termed, “MSI-69”) comprisingmicrocrystalline cellulose, croscarmellose, colloidal silicon dioxideand magnesium stearate was also generated using similar procedures asdescribed in Example 1. This formulation was directly compared at halfof the dose (800 mg) to a 1600 mg dose of the commercially availableS-adenosyl methionine tosylate disulfate formulation mentioned above.Seven healthy and fasted male volunteers were administered a single 800mg dose of MSI-69 and the presence of SAMe in plasma was measured afteradministration at the indicated time points.

As seen in the graph in FIG. 5C, the average C_(max) of the secondinventive formulation (MSI-69) when using only half the dose is alsocomparable to the full, 1600 mg dose of the commercially available SAMeformulation.

These results clearly show that the SAMe formulations of the inventionprovide a significantly reduced effective dose and thus improvedpharmacokinetic profile in comparison to approximately double the doseof the commercially available SAMe dosage forms used here.

A summary of the AUC and C_(max) values obtained from dosing SAMe at 800mg using either MSI-69 or MSI-72 as described above as well as twodifferent MSI formulations (MSI-90 and MSI-105) is shown in Table 4below in comparison to two commercially available SAMe products.

The compositions of the invention provide significantly higher exposurein comparison to the two commercially available SAMe products asmeasured by C_(max) and in particular, AUC at the 800 mg dose.

TABLE 4 Pharmacokinetic Analysis of 800 mg SAMe Doses AUC (ng · h/mL)Cmax¹ Test Article Dose 0-24 hours (ng/mL) Commercial 800 mg, BID  3572²784 SAMe #1 (t = 0, and 8 hrs) Commercial 800 mg, QD 3311 595 SAMe #2MSI-69 800 mg, QD 5279 931 MSI-72 800 mg, QD 4590 851 MSI-90 800 mg, QD5301 865 MSI-105 800 mg, QD 6372 1106 ¹The Cmax values are derived byaveraging the concentration of all subjects at each time point. ²The AUCdetermination for the Commercial SAMe product #1 was calculated byadjusting the SAMe plasma concentration for the 12 and 24 hour timepoint to the concentrations found in the pre-dose baseline level. Thisadjustment is necessary in order to remove the effects of the seconddose which was administered at the 8 hour time point, and thus allow anAUC comparison to the other periods which were only given a single SAMedose at t = 0.

Example 6 SAMe Formulations of the Invention Result in a Reduced FoodEffect Profile

Once a day dosing using the presented formulations was compared againsta simulated twice a day dosing of a routinely used commerciallyavailable SAMe product is provided. As mentioned previously,conventional SAMe formulations are typically dosed twice-daily (BID);and the second dose given late afternoon may contribute to insomniaand/or other sleep-related side effects (e.g. restlessness) because of a“food affect”, since the second dose is not administered to a fastedstomach.

The amount and/or type of food present in the stomach orgastrointestinal tract of an individual can cause a delay in the timetaken for an API within a tablet (or other dosage form) to be dissolvedand absorbed into the blood stream. In BID dosing of SAMe, the seconddose being administered to fed individuals is likely to delay the amountof time in which the second SAMe C_(max) will be achieved (i.e. a delayin the second T_(max)). Therefore, the C_(max) of the second dose (andpotential stimulatory effects associated with this) would occur duringnormal sleep hours which may explain why insomnia and othersleep-related side effects are common with conventional SAMe treatments.Once a day dosing using SAMe formulations of the invention, whichdeliver the equivalent total daily amount of SAMe as with conventionalbi-daily dosing, is likely to alleviate such side effects, particularlywhen administered in the early morning to fasted individuals, as onlyone C_(max) will occur daily and during normal waking hours.

FIG. 6 shows the “food affect” associated with a fed versus fastedadministration of a commercially available SAMe tosylate disulfateformulation, dosed twice-daily at 800 mg. The strong food affect is seenin the delay of T_(max) of the “fed” dosing to post 8 hours afteradministration of the single, 800 mg dose. This is in contrast to anaverage T_(max) of approximately four hours observed under fastedconditions. Clearly the presence of ingested food in the “fed” subjectscauses a significant delay in the time taken to reach the C_(max).

FIG. 7 represents a simulation of a 1600 mg bi-daily (BID) dosing of thecommercially available product given at time 0 hours fasted (8:00 am)and time 8 hours non-fasted or “fed” (4:00 pm). As seen in the graph,the first T_(max) is achieved within approximately 3-4 hours of thefirst dose given when the individuals had fasted. However, the secondT_(max), from the second dose given to “fed” individuals takes about 8hours to occur. The extended delay in T_(max) experienced in the seconddose under non-fasted conditions results in the C_(max) being reachedbetween 15 and 18 hours after the first dose and correlates to between11:00 pm to 2:00 am. This implies that the second dose is producing aSAMe C_(max) during normal sleep times, which may explain the presenceof insomnia and other sleep-related side effects associated with currentregimented BID dosing of SAMe.

Through the formulations exemplified in the present invention, once aday dosing has been shown to provide equivalent or higher SAMe AUCs withreduced T_(max) variability and a lack of high SAMe levels during normalnight sleeping times. The improved pharmacokinetic profile of theseformulations with once a day dosing should result in reduced side effectprofiles associated with SAMe administration.

A comparison of the simulated, conventional twice a day dosing regimento the formulations embodied in the present invention are shown below inTable 4:

TABLE 5 IN VIVO PROFILE (ranked by individual's averages) *Cmax ***AUCdose of (ng/mL **Tmax (ng h/mL SAMe ion of SAMe) (hrs) of SAMe)Simulated 1600 mg 2 × 800 mg     714 & 630 4 & 16  6445* BID commercialproduct MSI-79 1600 mg 2567 4 10250 MSI-72 1600 mg 2646 4 10365 MSI-781600 mg 3490 4 13642 MSI-77 1600 mg 3965 5 13927 MSI-43 1600 mg 4488 413168 1600 mg dose was given to subjects in one dose at 8am after anovernight fast *Average Cmax was calculated by averaging individual'sCmax values (no outliers) **Tmax is the time point where *Cmax wasdetermined ***AUC is area under the curve (AUC calculated for simulateddose by adding fed and fasted dosing profiles together)

1.-26. (canceled)
 27. A method for improving the pharmacokineticparameters of exogenous SAMe administered to a subject, said methodcomprising administering to the subject a non-parental compositioncomprising at least one physiologically effective dosage of SAMe incombination with at least one excipient selected from the groupconsisting of a matrix material, a binder, a lubricant, a glidant, acoating, a disintegrant, a super-disintegrant, a polysaccharide, anoligosaccharide, a polypeptide, a protein, a synthetic oligomer, asynthetic polymer, a monomeric organic molecule, a hydrophobic organicmolecule, a hydrophilic organic molecule, an amphoteric organicmolecule, an inorganic salt, an inorganic metal, and a combinationthereof, wherein said excipient improves the pharmacokinetic parametersof said SAMe in a subject, said pharmacokinetic parameters measurable inthe subject by one of a Cmax, an AUC, and combinations thereof incomparison to a selected SAMe reference data set.
 28. The method of toclaim 27, wherein the inorganic salts is a salt that provides an averageSAMe plasma area under the curve (average AUC) of at least about 7500ng·h/mL for a 1600 mg dosage of SAMe ion.
 29. (canceled)
 30. The methodof claim 27, wherein the composition is manufactured at a relativehumidity of less than 10%.
 31. The method of claim 27, wherein thecomposition is in a dosage form comprising a functional coating of from1 to 5% of the total weight of the dosage form.
 32. The method of claim31, wherein the composition when administered to a selected subjectgroup provides in the selected subject group an average C_(max) of atleast about 1800 ng/mL and an average SAMe plasma area under the curve(average AUC) of at least about 7500 ng·h/mL for a 1600 mg dosage ofSAMe ion.
 33. The method of claim 31, wherein the composition whenadministered to a selected subject group provides in the selectedsubject group an average C_(max) of at least about 850 ng/mL and anaverage SAMe plasma area under the curve (average AUC) of at least about4000 ng·h/mL for a 800 mg dosage of SAMe ion.
 34. The method of claim31, wherein the composition when administered to a selected subjectgroup provides in the selected subject group an average C_(max) of atleast about 100 ng/mL per 100 mg of SAMe ion in said physiologicallyeffective dosage.
 35. The method of claim 31, wherein the compositionwhen administered to a selected subject group provides in the selectedsubject group an average C_(max) within the range of about 100 ng/mL toabout 500 ng/mL per 100 mg of SAMe ion in said physiologically effectivedosage.
 36. The method of claim 31, wherein the composition whenadministered to a selected subject group provides in the selectedsubject group one of an average T_(max) with reduced variation or areduced effective dose in comparison to a SAMe reference data set. 37.The method of claim 31, wherein the composition when administered to aselected subject group provides in the selected subject group a reducedside effect profile in comparison to a SAMe reference data set.
 38. Themethod of claim 31, wherein the composition when administered to aselected subject group provides in the selected subject group an averageAUC of at least about 7500 ng·h/mL for a 1600 mg dosage of SAMe ion. 39.The method of claim 31, wherein the composition when administered to aselected subject group provides in the selected subject group an averageAUC of at least about 4000 ng·h/mL for a 800 mg dosage of SAMe ion. 40.The method of claim 31, wherein the composition when administered to aselected subject group provides in the selected subject group an averageAUC within the range of about 500 ng·h/mL to about 800 ng·h/mL per 100mg dosage of SAMe ion.
 41. A formulation comprising SAMe, wherein theformulation comprises a mixture of SAMe and at least one excipient andthe mixture is produced by combining said SAMe and said excipient at arelative humidity less than about 10%.
 42. A formulation comprisingSAMe, wherein the formulation comprises a mixture of SAMe and at leastone excipient, wherein the mixture exhibits a dissolution profile at pH6.0 suitable to target delivery to the proximal intestine.
 43. A processof improving the pharmacokinetic profile of a SAMe formulation,comprising manufacturing said SAMe formulation at a relative humidity ofless than about 10%.
 44. A composition for oral administration,comprising SAMe and at least one excipient wherein the formulationexhibits an in vitro dissolution profile in pH 6.0 aqueous solution suchthat greater than 20% and less than 90% of total SAMe in the compositionis dissolved from 30 to 90 minutes of incubation in said pH 6.0 aqueoussolution.
 45. The composition of claim 44, wherein the formulationexhibits an in vitro dissolution profile in pH 6.0 aqueous solution suchthat greater than 25% and less than 80% of total SAMe in the compositionis dissolved from 45 to 75 minutes of incubation in said pH 6.0 aqueoussolution.
 46. The composition of claim 44, wherein the composition is ina dosage form manufactured at a relative humidity of less than 10%. 47.The composition of claim 46, wherein the composition is in a dosage formthat comprises a functional coating and the functional coatingconstitutes less than or equal to 5% of the total weight of the dosageform.
 48. A method for improving the uptake of SAMe, comprisingadministering to a patient SAMe in a formulation that exhibits an invitro dissolution profile at pH 6.0, wherein greater than 20% and lessthan 90% of total SAMe is dissolved between 30 to 90 minutes ofincubation in said pH 6.0 aqueous solution.
 49. The method of claim 48,wherein the formulation exhibits an in vitro dissolution profile in pH6.0 aqueous solution such that greater than 25% and less than 80% oftotal SAMe in the composition is dissolved between 45 to 75 minutes ofincubation in said pH 6.0 aqueous solution.
 50. The method of claim 48,wherein the composition is in a unit dosage form manufactured at arelative humidity of less than 10%.
 51. The method of claim 48, whereinthe composition is in a unit dosage form that comprises a functionalcoating and the functional coating constitutes less than or equal to 5%of the total weight of the unit dosage form.
 52. The method of claim 48,wherein the composition is in a unit dosage form that comprises afunctional coating of from 1 to 5% of the total weight of the unitdosage form.
 53. The method of claim 37, wherein the reduced side effectprofile is measured as a reduction in gastrointestinal side effects,insomnia, restlessness, sexual dysfunction, lower homocysteine levels,cardiovascular side effects, nervousness, loss of appetite, dry mouth,dizziness or headaches.